FR2481712A1 - ENZYMATIC REDUCING AGENT FOR DETERGENT COMPOSITION - Google Patents

ENZYMATIC REDUCING AGENT FOR DETERGENT COMPOSITION Download PDF

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FR2481712A1
FR2481712A1 FR8108602A FR8108602A FR2481712A1 FR 2481712 A1 FR2481712 A1 FR 2481712A1 FR 8108602 A FR8108602 A FR 8108602A FR 8108602 A FR8108602 A FR 8108602A FR 2481712 A1 FR2481712 A1 FR 2481712A1
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cellulase
reducing agent
washing
hardness
detergent composition
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FR2481712B1 (en
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Peder Oe Barbesgaard
Georg Wilhelm Jensen
Poul Holm
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Novo Nordisk AS
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Novo Industri AS
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/24Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405Glucanases
    • C12N9/2434Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2437Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
    • CCHEMISTRY; METALLURGY
    • C11ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11DDETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16Organic compounds
    • C11D3/38Products with no well-defined composition, e.g. natural products
    • C11D3/386Preparations containing enzymes, e.g. protease or amylase
    • C11D3/38645Preparations containing enzymes, e.g. protease or amylase containing cellulase
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y302/00Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01004Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Detergent Compositions (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

L'INVENTION CONCERNE DES PERFECTIONNEMENTS RELATIFS A UN ADDITIF ENZYMATIQUE. L'ADDITIF ENZYMATIQUE REDUCTEUR DE DURETE DE L'INVENTION EST A BASE D'UNE CELLULASE FONGIQUE, C'EST-A-DIRE UNE CELLULASE PRODUITE AU MOYEN D'HUMICOLA INSOLENS. CET ADDITIF ENZYMATIQUE PEUT ETRE OBTENU AVEC DE HAUTS RENDEMENTS ET IL EXERCE UNE ACTIVITE EXTRAORDINAIREMENT PUISSANTE A DES PH ALCALINS. L'INVENTION EST UTILISEE POUR REDUIRE LA DURETE DES TISSUS SOUMIS A UN LAVAGE, DE MEME QUE POUR EFFECTUER UN LAVAGE EN UNE SEULE OPERATION.THE INVENTION CONCERNS IMPROVEMENTS RELATING TO AN ENZYMATIC ADDITIVE. THE HARDNESS-REDUCING ENZYMATIC ADDITIVE OF THE INVENTION IS BASED ON A FUNGAL CELLULASE, THAT IS, A CELLULASE PRODUCED BY MEANS OF HUMICOLA INSOLENS. THIS ENZYMATIC ADDITIVE CAN BE OBTAINED WITH HIGH YIELDS AND IT EXTRIES AN EXTRAORDINARILY POWERFUL ACTIVITY AT ALKALINE PH. THE INVENTION IS USED TO REDUCE THE HARDNESS OF FABRICS SUBJECT TO WASHING, AS WELL AS TO PERFORM A WASHING IN A SINGLE OPERATION.

Description

PERFECTICINNEiMENTS RELATIFS A UN ADDITIF ENZYlNATIQUE.PERFECTICINNEITIONS RELATING TO AN ENZYLNATIC ADDITIVE.

La présente invention concerne des perfec-  The present invention relates to

tionnements relatifs à un agent réducteur de dureté pour une composition détergente; l'invention concerne également une composition détergente et un procédé de lavage. On sait que le lavage de tissus contenant du coton dans des conditions normales est généralement la cause d'une dureté prononcée désagréable. Dans la  compositions relating to a hardness reducing agent for a detergent composition; the invention also relates to a detergent composition and a washing method. It is known that the washing of fabrics containing cotton under normal conditions is generally the cause of an unpleasant pronounced hardness. In the

pratique habituelle du lessivage, on réduit normale-  the usual practice of leaching, normal

ment cette dureté en traitant les tissus avec un agent  this hardness by treating the tissues with an agent

adoucissant (agent de conditionnement de rinçage) con-  softener (rinse conditioning agent)

tenant un détergent cationique et ce, au cours d'une étape distincte du lavage principal. Toutefois, ce traitement présente un sérieux inconvénient du fait que l'effet réducteur de dureté est d'une nature  holding a cationic detergent during a separate step of the main wash. However, this treatment has a serious disadvantage because the reducing effect of hardness is of a nature

temporaire. Après le lavage suivant, la dureté réap-  temporary. After the following wash, the hardness

paraî:t à moins que l'on ne traite à nouveau une fois de plus le tissu avec un agent adoucissant. Bien que d'importants investissements aient été consacrés en vue de trouver de meilleures méthodes pour réduire la dureté des tissus, il n'en reste pas moins vrai que les agents adoucissants utilisés dans la pratique ont  unless the fabric is again treated again with a softening agent. Although significant investments have been made to find better methods for reducing the hardness of the fabrics, it is still true that the softening agents used in practice have

dû être ajoutés aux tissus au cours d'une étape dis-  had to be added to the tissues during a step

tincte de l'étape de lavage principale pour exercer  tincte of the main wash step to exercise

pleinement leur effet réducteur de dureté; par ail-  fully reduce their hardness; otherwise

leurs, cet effet réducteur de dureté n'est que d'une  their, this reducing effect of hardness is only one

nature temporaire.temporary nature.

Toutefois, la littérature fait mention de  However, the literature mentions

différentes suggestions en vue de résoudre ce problè-  different suggestions for solving this problem.

me. Par exemple, dans la spécification du brevet britannique n0 1.368.599, on décrit, en variante, une méthode prétendument plus efficace pour réduire la dureté des tissus contenant du coton, cette méthode  me. For example, in British Patent Specification No. 1,368,599 there is alternatively described an allegedly more effective method for reducing the hardness of cotton-containing fabrics.

consistant à effectuer un traitement avec une cellu-  treatment with a cellulose

lase. Il est à noter que ce traitement est toujours effectué au cours d'une étape distincte, par exemple,  lase. It should be noted that this treatment is always performed during a separate step, for example,

248 1712248 1712

en guise de prélavage ou de prétrempage, ainsi qu'on l'exposera ci-après de manière plus détaillée mais, dans la spécification de ce brevet britannique nO 1.368.599, on prétend que les tissus contenant du coton et traités avec une cellulase conservent leur douceur au cours de plusieurs lavages. Toutefois, il est à noter que cet agent adoucissant à base de cellulase n'est pas apparu dans le commerce encore que cette technique fut connue dès 1972 (lors de la  as a pre-wash or pre-soak, as will be discussed below in more detail, but in the specification of this British Patent No. 1,368,599 it is claimed that cotton-containing fabrics treated with cellulase retain their softness during several washes. However, it should be noted that this softening agent based on cellulase did not appear commercially, although this technique was known as early as 1972 (when

publication du brevet belge no 773.280, qui est équi-  publication of Belgian Patent No. 773,280, which is equi-

valent au brevet britannique 1.368.599).  are British Patent 1,368,599).

Dès lors, la description donnée dans la spécification  Therefore, the description given in the specification

du brevet britannique n0 1.368.599 (ou brevet belge  British Patent No. 1,368,599 (or Belgian Patent

773.280) doit être considérée non comme un enseigne-  773.280) must be regarded not as a sign-

ment technique réel, mais bien comme une possibilité  really technical, but as a possibility

théorique que l'on envisage de commercialiser.  theoretical that we plan to market.

Il est à noter que les cellulases bactérien-  It should be noted that bacterial cellulases

nes sont habituellement assez coûteuses du fait que  are usually quite expensive because

leur production est normalement très faible.; les cel-  their production is normally very low .; the

lulases fongiques sont les plus prometteuses du point  fungal lulases are the most promising of the

de vue commercial car elles sont généralement produi-  from a commercial point of view as they are generally produced

tes avec des rendements un peu plus élevés. Par exem-  with a little higher yields. For example

ple, à la page 2, lignes 84-90 de la copie imprimée de la spécification du brevet britannique n0  on page 2, lines 84-90 of the printed copy of the specification of British Patent

1.368.599, il est dit: "De nombreux types de cellu-  1.368.599, it says: "Many types of cell

lases dérivant des champignons ont un pH optimum d'environ 5. Au-delà d'un pH de 7, leur activité est normalement fortement réduite et, par conséquent, les enzymes cellulolytiques dérivant des champignons doivent être utilisées dans un milieu acide suivant  The levels derived from fungi have an optimum pH of about 5. Above a pH of 7, their activity is normally greatly reduced and, therefore, cellulolytic enzymes derived from fungi must be used in a following acidic medium.

la présente invention". Ceci signifie que l'opéra-  the present invention. "This means that the operation

tion de lavage qui est normalement effectuée à un pH alcalin, ne peut avoir lieu au cours de la même étape que le traitement à la cellulase. On effectue ce traitement à la cellulase dans une première étape en guise de prélavage ou de prétrempage acide. après quoi on procède au lavage principal au cours d'une deuxième étape en guise de lavage alcalin. A la connaissance de la Demanderesse, la technique ne fait  This washing procedure, which is normally carried out at an alkaline pH, can not take place during the same step as the cellulase treatment. This cellulase treatment is carried out in a first step as a pre-wash or acid pre-soak. after which the main wash is carried out in a second step as an alkaline wash. To the knowledge of the Applicant, the technique does not

mention d'aucune description d'une cellulase devant  mention of no description of a cellulase in front of

S être utilisée pour effectuer un lavage et un adoucis-  To be used for washing and softening

sement en une seule étape au cours d'un procédé de  in a single step during a

lavage pratique.practical washing.

Comme on le comprendra, du fait qu'il est nécessaire dieffectuer le traitement à la cellulase à un pH acide avant le lavage principal, on doit  As will be understood, since it is necessary to perform the cellulase treatment at an acidic pH before the main wash, one must

procéder à un rinçage intermédiaire entre le traite-  carry out an intermediate rinsing between

ment acide à la cellulase et le lavage principal à moins qu'un excès d'un alcali destiné à neutraliser l'acidité de la lessive traitée à la cellulase ne soit présent dans le détergent destiné au lavage principal ou à moins que cet excès dtalcali ne soit  cellulase acid and the main wash unless an excess of an alkali to neutralize the acidity of the cellulase-treated detergent is present in the detergent for the main wash or unless the excess is

ajouté à ce détergent.added to this detergent.

Enfin, si le liquide de traitement à la cel-  Finally, if the treatment liquid with

lulase contient des détergents, l'aptitude au nettoy-  lulase contains detergents, cleanability

age du détergent est normalement assez médiocre aux  the detergent is normally quite poor

faibles pH dans le liquide de traitement à la cellula-  low pH in the cellulosic treatment liquid.

se. Dès lors, il est urgent de trouver, pour un lavage principal, une composition détergente contenant une cellulase relativement bon marché et exerçant une haute activité aux pH régnant normalement dans les  is. Therefore, it is urgent to find, for a main wash, a detergent composition containing a relatively inexpensive cellulase and exerting a high activity at the pH levels normally prevailing in the

solutions de lavage principal.main wash solutions.

Toutefois, la technique ne fait mention d'au-  However, the technique does not mention

cune cellulase économique ayant une haute activité  an economic cellulase with high activity

acceptable aux pH régnant normalement dans les solu-  acceptable at the pH levels normally found in

tions de lavage principal en dépit du fait que l'on pourrait statendre à réaliser un profit commercial  main washers despite the fact that commercial profit could be expected to

remarquable si ce besoin pouvait être satisfait.  remarkable if this need could be satisfied.

A présent, de façon étonnante, suivant l'in-  Now, surprisingly, following the in-

vention, on a trouvé qu'une certaine-cellulase fongi-  vention, some fungal cellulase has been found to be

que, c'est-à-dire la cellulase fongique pouvant 8tre  that is, fungal cellulase which can be

produite à partir d'Humicola insolens (Humicola gri-  produced from Humicola insolens (Humicola

sea var. thermoidea), exerçait une haute activité  sea var. thermoidea), exercised a high activity

de cellulase aux pH régnant normalement dans les so-  cellulase at the pH levels normally prevailing in

lutions de lavage principal et ce, contrairement à ce que l'on pouvait escompter sur la base des connais- sances de la technique comme indiqué, par exemple, dans le passage précité de la spécification du brevet  the main washings, contrary to what could be expected on the basis of the knowledge of the art as indicated, for example, in the aforementioned passage of the patent specification.

britannique n 1.368.599.British 1,368,599.

Dès lors, suivant le premier aspect de la pré-  Therefore, according to the first aspect of the pre-

sente invention, on prévoit un agent réducteur de du-  In this invention, a reducing agent of

reté sous forme d'un additif pour une composition détergente, un composant essentiel de cet agent étant une cellulase fongique pouvant être produite à partir  in the form of an additive for a detergent composition, an essential component of this agent being a fungal cellulase that can be produced from

d'une souche d'Humicola insolens (Humicola grisea var.  of a strain of Humicola insolens (Humicola grisea var.

thermoidea), cet agent réducteur de dureté étant des-  thermoidea), this hardness reducing agent being

tiné à une composition détergente pour un lavage prin-  detergent composition for a main wash

cipal. En d'autres mots, l'invention consiste à utiliser la cellulase fongique pouvant être produite à partir d'Humicola insolens comme agent réducteur de dureté pour une composition détergente destinée à un  cipal. In other words, the invention is to use the fungal cellulase that can be produced from Humicola insolens as a hardness reducing agent for a detergent composition intended for

lavage principal.main wash.

Il est entendu que la cellulase fongique pouvant être obtenue à partir d'Hlumicola insolens peut être produite à partir de souches ordinaires d'Humicola insolens, ainsi qu'à partir de mutants  It is understood that the fungal cellulase obtainable from Hlumicola insolens can be produced from ordinary strains of Humicola insolens, as well as from mutants

et de variants d'Humicola insolens.and variants of Humicola insolens.

En utilisant l'agent réducteur de dureté pour une composition détergente destinée à un lavage principal conformément à l'invention, il est devenu  By using the hardness reducing agent for a detergent composition for a main wash according to the invention, it has become

possible pour la première fois, d'effectuer un procé-  possible for the first time, to carry out a procedure

dé de lavage et de réduction de dureté d'une manière raisonnablement économique et en une seule opération,  washing and hardness reduction in a reasonably economical manner and in a single operation,

c'est-à-dire au cours du lavage principal, sans de-  that is to say during the main wash, without

voir recourir à un prétrempage ou à un autre traite-  see pre-soaking or other treatment

ment préalable.beforehand.

Dans "Agric. Biol. Cheim." 44 (3), 481-487 (1980) et 44 (8), 1721-1728 (1980), il est stipulé que l'Humicola insolens est un champignon thermiiophile producteur de cellulase. De même, il est stipulé que les pH optima des cellulases produites à partir  In "Agric Biol Cheim." 44 (3), 481-487 (1980) and 44 (8), 1721-1728 (1980), it is stated that Humicola insolens is a thermophilic cellulase-producing fungus. Similarly, it is stipulated that the optimum pH of cellulases produced from

dtHumicola insolens sont de 5,3 et de 5,0 respective-  dtHumicola insolens are 5.3 and 5.0 respectively.

ment, si bien que cette cellulase semblerait apparte-  so that this cellulase would seem to belong

nir à la catégorie des cellulases indiquées ci-dessus à propos du brevet britannique n 1.368.599; dès lors, il semblerait que ces cellulases soient totalement inaptes à réaliser l'objet de l'invention. Toutefois, de façon étonnante, suivant l'invention, on a trouvé  to the category of cellulases mentioned above in relation to British Patent No. 1,368,599; therefore, it would appear that these cellulases are totally unfit to achieve the object of the invention. However, surprisingly, according to the invention, it has been found

que Humicola insolens produisait une cellulase réali-  that Humicola insolens was producing a cellulase

sant pleinement l'objet de l'invention.  fully the object of the invention.

Humicola insolens rentre dans le groupe de la souche thermophile Humicola, laquelle comprend Humicola insolens et Humicola var. thermoidea, la distinction taxonomique entre ces deux souches étant  Humicola insolens belongs to the group of the thermophilic strain Humicola, which includes Humicola insolens and Humicola var. thermoidea, the taxonomic distinction between these two strains being

très douteuse.very doubtful.

L'activité de cellulase régulière Cx est  Cx regular cellulase activity is

déterminée en raison du fait que la cellulase hydro-  determined by the fact that hydrous cellulase

lyse la carboxyméthyl-cellulose en carbohydrates  lysis carboxymethyl cellulose to carbohydrates

réducteurs dont l'aptitude à la réduction est déter-  reducing agents whose ability to reduce

minée par colorimétrie par réaction au ferricyanure comme décrit par Hoffman, W.S., "J. Biol. Chem." 120,  by colorimetry by reaction with ferricyanide as described by Hoffman, W.S., J. Biol Chem. 120,

51 (1937).51 (1937).

L'activité de cellulase régulière C est x déterminée par rapport à la carboxyméthyl-cellulose  The regular cellulase activity C is x determined relative to carboxymethylcellulose

constituant le substrat ("tCMC Hercules, 4M6F").  constituting the substrate ("tCMC Hercules, 4M6F").

Les conditions classiques sont les suivantes: pH = 9,50; tampon: tris(hydroxyméthyl)aminométhane  The standard conditions are as follows: pH = 9.50; buffer: tris (hydroxymethyl) aminomethane

("tris") 0,1M; substrat: 4 g de carboxyméthyl-  ("sorting") 0.1M; Substrate: 4 g carboxymethyl

cellulose/litre de la solution tampon indiquée ci-  cellulose / liter of the buffer solution indicated

dessus; température d'incubation: 50 C; temps  above; incubation temperature: 50 C; time

d'incubation = 20 minutes.incubation time = 20 minutes.

Une unité d'activité (le cellulase régulière Cx (que l'on appellera plus brièvement ci-après "une unité d'activité réglièrede Cx") est la quantité de cellulase qui, dans les conditions classiques indiquées ci-dessus, forme un sucre réducteur en une quantité équivalant  One unit of activity (Cx regular cellulase (which will be referred to more briefly hereinafter as "a unit of Cx ruled activity") is the amount of cellulase which, under the conventional conditions indicated above, forms a sugar. reducer in an equivalent amount

à 1 rmole de glucose par minute.at 1 mole of glucose per minute.

Dans une forme de réalisation préférée de l'agent réducteur de dureté suivant l'invention, la cellulase fongique est produite au moyen de la souche  In a preferred embodiment of the hardness reducing agent according to the invention, the fungal cellulase is produced by means of the strain

Humicola DSM 1800. On a trouvé que cette souche pro-  Humicola DSM 1800. It has been found that this strain

duisait une cellulase alcaline ayant une haute activité de C à des pH alcalins. La nouvelle souche Humicola  alkaline cellulase with a high activity of C at alkaline pH. The new strain Humicola

a été identifiée au "Commonwealth Mycological Insti-  identified in the Commonwealth Mycological Insti-

tute", Kew, Grande-Bretagne, sous le nom "Humicola insolens". Cette nouvelle souche a été déposée le  tute ", Kew, Great Britain, under the name" Humicola insolens "This new strain was deposited on

14 avril 1980 à la "German Collection of Microorga-  14 April 1980 to the "German Collection of Micro-

nisms" (DSM, "Deutsche Sammlung von Mikroorganismen"), G6ttingen, République Fédérale d'Allemagne, sous le  nisms "(DSM," Deutsche Sammlung von Mikroorganismen "), G6ttingen, Federal Republic of Germany, under the

n0 DSM 1800.n0 DSM 1800.

On donnera ci-après une description morpholo-  A morphological description will be given below.

gique et physiologique de la nouvelle souche Humicola  and physiology of the new strain Humicola

DSM 1800 mentionnée ci-dessus.DSM 1800 mentioned above.

Croissance: courte; mycélium tout d'abord blanc, de-  Growth: short; mycelium first of all white,

venantplus tard gris foncé avec des taches blanches;  coming later dark gray with white spots;

hyphes incolores, lisses, cloison d'un diamètre d'en-  colorless, smooth hyphae, septum with a diameter of

viron 2,8 ô. Gouttes d'eau incolores parfois pré-  viron 2,8 ô. Colorless water drops sometimes pre-

sentes.paths.

Réverse: jaune devenant brun/gris foncé avec l'âge.  Reverse: yellow becoming brown / dark gray with age.

Aleuriospores: lisses; brunes; produites indivi-  Aleuriospores: smooth; brown; produced individually

duellement, en terminaison de conidiophores indivi-  dually, in termination of individual conidiophores

duels ou multicellulaires; habituellement globeuses  dual or multicellular; usually globose

12,6-16,8ï ou parfois subglobeuses 7 x 11,2 -  12.6-16.8 or sometimes subglobose 7 x 11.2 -

14 x 16,8. Apicule présent.14 x 16.8. Apicle present.

Chlamydospores: produites individuellement ou en  Chlamydospores: produced individually or in

chaîne; lisses, brunes; 11,2-16,8 ô.  chain; smooth, brown; 11,2-16,8 ô.

Phialocondies: absentes.Phialocondias: absent.

Limites de température: pas de croissance à 260C; bonne croissance à partir de 37-50 C. Colonies de  Temperature limits: no growth at 260C; good growth from 37-50 C. Colonies of

8 cm de diamètre à 37 C pendant 5 jours.  8 cm in diameter at 37 C for 5 days.

Les observations ci-dessus ont été relevées après 5 jours à 37 C sur un milieu d'YPSS/agar-agar ayant la composition indiquée ci-dessous: Extrait de levure Difco 4 g K2HPO4 1 g MgSO4.7H20 0,5 g Amidon soluble 15 g Eau distillée 1.000 ml  The above observations were recorded after 5 days at 37 ° C. on a YPSS / agar medium having the composition indicated below: Difco yeast extract 4 g K 2 HPO 4 1 g MgSO 4 .7H 2 O 0.5 g Soluble starch 15 g distilled water 1,000 ml

Agar-agar 15 g.Agar-agar 15 g.

De façon étonnante, on a trouvé que, en plus de son effet adoucissant, la cellulase produite au  Surprisingly, it has been found that, in addition to its softening effect, cellulase produced at

moyen de la souche Humicola insolens DSM 1800 exer-  medium strain Humicola insolens DSM 1800 exert

çait un puissant effet de détachement de la saleté et d'anti-redéposition. Dans une forme de réalisation préférée de l'agent réducteur de dureté suivant l'invention, la  had a strong detaching effect from dirt and anti-redeposition. In a preferred embodiment of the hardness reducing agent according to the invention, the

cellulase fongique est enrichie vis-à-vis de la frac-  fungal cellulase is enriched with respect to

tion de cellulase qui ne se fixe pas à un échangeur  cellulase that does not attach to a heat exchanger

d'anions à 6,5 L pH ú 7,5.anion at 6.5 L pH ú 7.5.

Comme on l'indiquera en détail ci-après  As will be detailed below

(voir exemple 6), on a trouvé que la fraction de cel-  (see Example 6), it has been found that the fraction of this

lulase ne se fixant pas à un échangeur d'anions à  lulase does not bind to an anion exchanger at

6,5 t pH ' 7,5 (que l'on appellera ci-après plus bri-  6.5 t pH '7.5 (hereinafter referred to as more

èvement "fraction ACxIl') possède l'aptitude la plus significative à réduire la dureté. On a également constaté que la stabilité thermique de la fraction "ACxIu' était supérieure à celle des autres fractions de la cellulase dtHumicola insolens, ainsi qu'à celle  The "ACxIl fraction" has the most significant ability to reduce hardness, and the thermal stability of the "ACxIu" fraction has been found to be superior to that of the other cellulase fractions of Humicola insolens, as well as

des cellulases fongiques acides décrites dans le bre-  acid fungal cellulases described in the patent

vet britannique précité n 1.368.559.  aforesaid British veil No. 1,368,559.

Afin de pouvoir établir la teneur de la  In order to establish the content of the

fraction "ACx I", on peut déterminer l'activité modi-  "ACx I" fraction, it is possible to determine the

248 1712248 1712

fiée deCx (cellulase) de la manière décrite ci-après.  Cx (cellulase) as described hereinafter.

Un tampon de phosphate A est constitué de 26,52 g de dihydroxy-phosphate de sodium à deux molécules d'eau dans 1.000 ml d'eau déminéralisée; dès lors, la molarité de A est de 0,17. On mélange 100 ml de A avec environ 800 ml d'eau déminéralisée, on règle le pH à 7 et on règle le volume total du tampon à 1.000  A phosphate buffer A consists of 26.52 g of sodium dihydroxyphosphate with two molecules of water in 1,000 ml of demineralized water; therefore, the molarity of A is 0.17. 100 ml of A are mixed with approximately 800 ml of demineralized water, the pH is adjusted to 7 and the total volume of the buffer is adjusted to 1,000.

ml (appelé tampon B) dont la molarité est de 0,017.  ml (called buffer B) whose molarity is 0.017.

Dans une colonne, on charge 1 g de l'échangeur d'anions "DEAE-Sepharose C1-6B" (marque commerciale  In a column, charge 1 g of the anion exchanger "DEAE-Sepharose C1-6B" (trademark

déposée) et on l'équilibre avec le tampon B. On dis-  deposited) and equilibrated with buffer B.

sout ensuite l'enzyme dans le tampon B et on fait  then support the enzyme in buffer B and make

passer 25 ml de la solution enzymatique ainsi prépa-  25 ml of the enzymatic solution thus prepared

rée à travers la colonne équilibrée indiquée ci-des-  through the balanced column indicated below.

sus. On détermine l'activitédeC de la solution enzy-  extra. The C activity of the enzyme solution is determined

matique en aval de la colonne de la manière indiquée  downstream of the column as shown

ci-dessus pour la détermination de l'activité réguliè-  above for the determination of the regular activity

re de cellulase Cx. Cette activité de Cx est appelée  cellulase Cx. This activity of Cx is called

"activité modifiéede C " et elle est inférieure à l'ac-  "modifiedC activity" and is lower than the

x tivité régulière de CÀ lorsqu'une certaine activité de C  Regularity of C when a certain activity of C

est adsorbée sur l'échangeur d'anions dans les condi-  is adsorbed on the anion exchanger under the conditions

tions indiquées.indicated.

Dans une forme de réalisation préférée de l'invention, la cellulase fongique présente un rapport (activité modifiée de CX/activité régulière de C) d'au  In a preferred embodiment of the invention, the fungal cellulase has a ratio (modified activity of CX / regular activity of C) of

moins 0,6, de préférence, d'au moins 0,8.  minus 0.6, preferably at least 0.8.

On a constaté qu'un agent réducteur de dure-  It has been found that a reducing agent

té de ce type était enrichi vis-à-vis de la fraction "'ACxI"' à un point tel que l'aptitude à la réduction x  This type was enriched with respect to the "ACxI" fraction to such an extent that the reduction ability

de la dureté est extraordinairement bonne.  Hardness is extraordinarily good.

Dans une forme de réalisation préférée de l'agent réducteur de dureté suivant l'invention, la cellulase fongique est prévue sous forme de granulés ne dégageant pas de poussières. Il est essentiel que la préparation de cellulase ait une faible teneur en poussières. En présence d'une importante quantité de poussières, des difficultés peuvent surgir à la fois dans les usines o la préparation de cellulase est mélangée avec des détergents, ainsi qu'au cours du traitement ultérieur du détergent contenant la cellulase. On peut obtenir une préparation de cellu- lase en granulés de diverses manières, par exemple, au moyen d'un appareil appelé IlMarumerizer" (= marque commerciale) comme décrit dans les spécifications des brevets britanniques n0 1.362.365 et 1.361.387 ou au moyen d'une machine de formation de granulés du type décrit dans "Aufbereitungstechiiikt' n0 3/1970, pages 147-153 et n0 5/1970, pages 262278; de même, cette préparation peut se présenter sous forme de perles comme décrit dans la spécification du brevet belge n0 760.135. Toutefois, dans tous les cas, les granulés doivent avoir des propriétés donnant lieu à un faible  In a preferred embodiment of the hardness reducing agent according to the invention, the fungal cellulase is provided in the form of granules which do not give off dust. It is essential that the cellulase preparation has a low dust content. In the presence of a large amount of dust, difficulties can arise both in the factories where the cellulase preparation is mixed with detergents, as well as during the subsequent treatment of the detergent containing the cellulase. A granular cellulase preparation can be obtained in various ways, for example, by means of a device called IlMarumerizer ™ (= trade mark) as described in British Patent Specification Nos. 1,362,365 and 1,361,387 or US Pat. by means of a granulation machine of the type described in "Aufbereitungstechiiikt" No. 3/1970, pages 147-153 and 5/1970, pages 262278; likewise, this preparation may be in the form of beads as described in the specification of Belgian Patent No. 760,135. However, in all cases, the granules must have properties giving rise to a low

dégagement de poussières.release of dust.

Dans une forme de réalisation préférée de l'agent réducteur de dureté suivant l'invention, les granulés de cellulase sont enrobés d'un agent de blanchiment, de préférence, TiO2, en combinaison avec  In a preferred embodiment of the hardness reducing agent according to the invention, the cellulase granules are coated with a bleaching agent, preferably TiO 2, in combination with

un agent fixant les poussières. De la sorte, on ob-  an agent fixing the dust. In this way, we ob-

tient des granulés de cellulase d'un bel aspect déga-  holds cellulase granules with a beautiful appearance

geant un minimum de poussières.with a minimum of dust.

Selon une autre forme de réalisation préfé-  According to another preferred embodiment

rée de l'agent réducteur de dureté suivant l'inven-  of the hardness reducing agent according to the invention

tion, ce dernier est un liquide dans lequel la cellu-  tion, the latter is a liquid in which the cell is

lase fongique est prévue sous forme d'un concentrat  fungal lase is expected in the form of a concentrate

en suspension dans un agent tensio-actif non ionique.  suspended in a nonionic surfactant.

Le concentrat est obtenu sur la base du bouillon de fermentation. Ainsi qu'on lta mentionné précédemment,  The concentrate is obtained on the basis of the fermentation broth. As mentioned previously,

il est essentiel que la préparation de cellulase dé-  it is essential that the cellulase preparation de-

gage peu de poussières. Etant donné que, dans cette forme de réalisation, l'agent réducteur de dureté  pledge little dust. Since in this embodiment the hardness reducing agent

suivant l'invention est un liquide, sa teneur en pous-  according to the invention is a liquid, its content in pus-

sières est nulle et, dès lors, on évite totalement les difficultés précitées. Lorsque l'agent réducteur de dureté est une suspension, l1agent tensio-actif non  is void and, therefore, the above difficulties are avoided altogether. When the hardness reducing agent is a suspension, the nonionic surfactant is

ionique contient avantageusement un agent épaissis-  ionic composition advantageously contains a thickening agent

sant. De la sorte, on peut normalement éviter la sédimentation de la cellulase. Comme agent épaissis-  health. In this way, the sedimentation of cellulase can normally be avoided. As a thickening agent

sant, on peut utiliser, par exemple, de la silice py-  However, for example, silica py-

rogénée en particules d'une granularité extrêmement faible. Si lton utilise de la silice pyrogénée comme  rogenous particles of extremely low granularity. If lton uses fumed silica as

agent épaississant, sa quantité se situe habituelle-  thickening agent, its quantity is usually

ment dans l'intervalle allant de 0,5 à 10% en poids/ poids, de préférence, dans 1lintervalle allant de  within the range of 0.5 to 10% w / w, preferably in the range from

1 à 5% en poids/poids.1 to 5% w / w.

Selon une forme de réalisation préférée de l'agent réducteur de dureté suivant l'invention, ce dernier est un liquide et la cellulase- fongique est prévue sous forme d'un concentrat de cellulase dissous dans un milieu aqueux, de préférence, en présence d'un agent stabilisant. Comme agent stabilisant, on peut  According to a preferred embodiment of the hardness reducing agent according to the invention, the latter is a liquid and the fungal cellulase is provided in the form of a cellulase concentrate dissolved in an aqueous medium, preferably in the presence of a stabilizing agent. As a stabilizing agent, one can

utiliser n'importe quel agent assurant une stabilisa-  use any agent providing stabilization

tion contre la sédimentation et contre la perte de  against sedimentation and against the loss of

l'activité enzymatique.enzymatic activity.

Suivant le deuxième aspect de la présente invention, on prévoit une composition détergente de lavage principal comprenant des ingrédients détergents et l'agent réducteur de dureté constituant le premier  According to the second aspect of the present invention, there is provided a main wash detergent composition comprising detergent ingredients and the hardness reducing agent constituting the first

aspect de la présente invention en une quantité per-  aspect of the present invention in a

mettant de réduire la dureté de telle sorte que le pH d'une solution de 1 g de la composition détergente de lavage principal dans un litre d'eau d'une dureté de 100 selon les normes allemandes avant l'addition de cette composition détergente de lavage principal, soit compris entre 7 et 10, de préférence, entre 7,5  reducing the hardness so that the pH of a solution of 1 g of the main wash detergent composition in one liter of water of a hardness of 100 according to German standards prior to the addition of this detergent composition of main wash, between 7 and 10, preferably between 7.5

et 9,5.and 9.5.

Il est entendu que l'expression "composition  It is understood that the expression "composition

détergente de lavage principal" désigne une composi-  washing detergent "means a composition

tion détergente qui est utilisée habituellement, mais  detergent which is usually used but

248 1712248 1712

non nécessairement, au cours d'un seul lavage qui ne doit pas être précédé ou suivi d'autres traitements de la nature d'un lessivage. L'avantage qu'offre la composition détergente de lavage principal suivant la présente invention apparalt très clairement lorsqu'- elle est utilisée au cours d'un seul lavage qui n'est  not necessarily, during a single wash that should not be preceded or followed by other treatments of the nature of leaching. The advantage of the main wash detergent composition according to the present invention becomes very apparent when it is used in a single wash which is not

pas précédé ou suivi d'autres traitements de 'la natu-  not preceded or followed by other treatments of 'natural

re d'un lessivage.re leaching.

Les ingrédients détergents d'une composition détergente de lavage principal suivant la présente invention ne sont pas critiques, la seule limitation qui leur est imposée, étant leur compatibilité -avec  The detergent ingredients of a main wash detergent composition according to the present invention are not critical, the only limitation imposed on them being their compatibility -with

la cellulase fongique.fungal cellulase.

A titre d'exemple, les ingrédients déter-  For example, the ingredients

gents et les pourcentages dans lesquels ils sont uti-  and the percentages in which they are used.

lisés, peuvent être les suivants: 1) Des agents tensio-actifs, en particulier, agents tensio-actifs anioniques et non ioniques, en  can be: 1) Surfactants, in particular anionic and nonionic surfactants, in particular

une quantité totale de 1 à 100% en poids, spécifique-  a total amount of 1 to 100% by weight, specifically

ment, en une quantité de 5 à 45%. Parmi les agents tensio-actifs anioniques spécifiques, il y a les alkyl-aryl-sulfonates linéaires et les sulfonates dia-oléfines. Des agents tensio-actifs non ioniques spécifiques sont les alkyl-phényl-éthoxylates et les  in an amount of 5 to 45%. Among the specific anionic surfactants are linear alkyl aryl sulfonates and diaolefin sulphonates. Specific nonionic surfactants are alkyl phenyl ethoxylates and

éthoxylates d'alcools gras.fatty alcohol ethoxylates.

2) Des charges, en particulier, des charges alcalines et des agents réduisant la dureté de l'eau,  2) Charges, in particular, alkaline fillers and agents reducing the hardness of water,

en une quantité totale de 5 à 80% en poids, spécifi-  in a total amount of 5 to 80% by weight, specifically

quement, de 25 à 75%. Des charges spécifiques sont  from 25 to 75%. Specific charges are

le tripolyphosphate de sodium, les silicates de so-  sodium tripolyphosphate, sodium silicates

dium-aluminium (zéolites), les silicates de sodium  dium-aluminum (zeolites), sodium silicates

et les carbonates de sodium.and sodium carbonates.

3) Des agents de blanchiment, en particulier, des peroxydes, en une quantité totale de 0 à 40% en poids, spécifiquement, de 0% ou de 15 à 30%. Des agents de blanchiment spécifiques sont le perborate  3) Bleaching agents, in particular peroxides, in a total amount of 0 to 40% by weight, specifically 0% or 15 to 30%. Specific bleaching agents are perborate

de sodium et le percarbonate de sodium.  of sodium and sodium percarbonate.

4) Dtautres ingrédients, principalement des enzymes, des agents d'avivage optique, des parfums,  4) Other ingredients, mainly enzymes, optical brighteners, perfumes,

des colorants, des modificateurs de mousses, des sta-  dyes, foaming modifiers, stain

bilisants, des agents anti-redéposition (mais, de préférence, non la carboxyméthyl-cellulose qui, dans une certaine mesure, est décomposée par la cellulase)  bilisants, anti-redeposition agents (but, preferably, not carboxymethyl cellulose which to some extent is decomposed by cellulase)

en une quantité totale de O à 10% en poids, spécifi-  in a total amount of from 0 to 10% by weight, specifically

quement de 0,1 à 5%.only 0.1 to 5%.

5) Des charges, en particulier, le sulfate de sodium et l'eau, en une quantité calculée de façon à compléter la formulation détergente à un total de %. Dans la brochure intitulée 'Novo Enzymes"  5) Fillers, in particular sodium sulphate and water, in an amount calculated to complete the detergent formulation to a total of%. In the brochure titled 'Novo Enzymes'

pour l'industrie des détergents (B 157a-GB 2000, fé-  for the detergent industry (B 157a-GB 2000,

vrier 1977), on donne une liste d'autres exemples généraux de formules détergentes que l'on pourrait  February 1977), a list of other general examples of detergent formulations that could be

utiliser conjointement avec la préparation de cellu-  use together with the cell preparation

lase, notamment des compositions caractérisées par  in particular compositions characterized by

des composés ou des catégories de composés spécifi-  compounds or classes of specific compounds

ques, de même que leurs pourcentage correspondants ou  as well as their corresponding percentage or

leurs intervalles de pourcentages correspondants.  their corresponding percentage intervals.

A la page 5 de la demande de brevet danois n 5691/78 déposée le 12 décembre 1978, on donne une  On page 5 of the Danish patent application No. 5691/78 filed December 12, 1978, a

liste de certaines compositions détergentes spécifi-  list of certain specific detergent compositions

ques que lion pourrait utiliser conjointement avec la  which could be used jointly with the

préparation de cellulase.cellulase preparation.

Dans une forme de réalisation préférée de la composition détergente de lavage principal suivant la présente invention, l'agent réducteur de dureté est présent en une quantité correspondant à 2,5-100 unités d'activité régulière de Cx/g du détergent de  In a preferred embodiment of the main wash detergent composition according to the present invention, the hardness reducing agent is present in an amount corresponding to 2.5-100 units of regular activity of Cx / g of the detergent of

lavage principal ou à 1,5-60 unités d'activité modi-  main wash or at 1.5-60 units of modified activity

fiée de Cx/g du détergent de lavage principal, de préférence, à 5-50 unités d'activité régulière de Cx/g dudétergent de lavage principal ou à 3-30 unités  Cx / g of main wash detergent, preferably 5-50 units of regular Cx / g activity of main wash detergent or 3-30 units

*2481712 2* 2481712 2

d'activité modifiée de Cx/g de détergent de lavage principal. De la sorte, on peut obtenir un effet  modified activity of Cx / g of main wash detergent. In this way, we can obtain an effect

réducteur de dureté satisfaisant et llon peut égale-  reducing hardness satisfactory and llon can also

ment éviter un excès de cellulase (qui rendrait la composition détergente de lavage principal non écono- mique). Selon une forme de réalisation préférée dc la composition détergente de lavage principal suivant  avoid excess cellulase (which would make the main wash detergent composition uneconomic). In accordance with a preferred embodiment of the following main wash detergent composition

la présente invention, cette dernière contient égale-  the present invention, the latter also contains

ment une protéinase bactérienne, de préférence, une protéinase obtenue au moyen de Bacillus lichenformis, en particulier, au moyen d"IALCALASE". De façon étonnante, on a trouvé que les additifs détergents,  Bacterial proteinase, preferably a proteinase obtained by means of Bacillus lichenformis, in particular, by means of "IALCALASE". Surprisingly, it has been found that detergent additives,

à savoir la cellulase et la protéinase, étaient com-  namely cellulase and proteinase, were

patibles et exerçaient chacun un effet.satisfaisant aux pH régnant au cours du procédé de lavage, bien que l'on puisse penser que l'enzyme protéolytique digère la cellulase. Cet additif détergent combiné exerce à la fois un meilleur effet de nettoyage et un meilleur effet adoucissant lorsqu'il est ajouté au détergent. L'expression "IALCALASE"II est une marque commerciale pour une préparation enzymatique vendue  and each had a satisfactory pH-effect during the washing process, although it is believed that the proteolytic enzyme digests cellulase. This combined detergent additive has both a better cleaning effect and a better softening effect when added to the detergent. The term "IALCALASE" II is a trademark for a sold enzyme preparation

dans le commerce par la firme "Novo". Si l'on utili-  in the trade by the firm "Novo". If we use

se lt"ALCALASE"I et un granulé de cellulase, on peut  lt "ALCALASE" I and a cellulase granule, one can

préparer l'additif enzymatique mixte soit en mélan-  prepare the mixed enzymatic additive either in

geant un granulé préalablement préparé d"'ALCALASE" avec un granulé préalablement préparé de cellulase, soit en m1élangeant un concentrat d' "ALCALASE"I avec un  previously prepared granule of "ALCALASE" with a previously prepared cellulase granule, or by mixing an "ALCALASE" concentrate with a

concentrat de cellulase, puis en introduisant ce mé-  cellulase concentrate and then introducing this

lange dans un dispositif de granulation, conjointe-  in a granulation device, together with

ment avec les produits auxiliaires habituels de granu-  with the usual auxiliary products of granular

lation. Selon une forme de réalisation préférée de la  lation. According to a preferred embodiment of the

composition détergente de lavage principal de la pré-  washing detergent composition of the present invention.

sente invention, un perborate est un des ingrédients détergents. Dans certaines conditions, ce perborate  In this invention, a perborate is one of the detergent ingredients. Under certain conditions, this perborate

248 1712248 1712

assure une meilleure efficacité de lavage et, de façon étonnante, on a trouvé que le perborate et la cellulase intervenant dans la composition détergente  assures a better washing efficiency and, surprisingly, it has been found that perborate and cellulase involved in the detergent composition

de lavage principal suivant l'invention étaient compa-  main wash according to the invention were

tibles. Suivant le troisième aspect de la présente invention, on prévoit un procédé de lavage principal, procédé dans lequel on utilise, comme détergent, une  patible. According to the third aspect of the present invention, there is provided a main washing process, wherein the detergent is

composition détergente de lavage principal constitu-  washing detergent composition

ant le deuxième aspect de la présente invention.  the second aspect of the present invention.

Selon une forme de riéalisation préférée de  According to a preferred embodiment of

ce procédé de lavage principal de la présente inven-  this main washing process of the present invention

tion, on utilise la cellulase fongique dans la solu-  fungal cellulase is used in the solution.

tion de lavage en une concentration correspondant à  washing concentration in a concentration corresponding to

10-100 unités d'activité régulière de Cx/litre de so-  10-100 regular activity units of Cx / liter of

lution de lavage ou à 6-60 unités d'activité modifiée de CX/litre de solution de lavage, de préférence, à  washing solution or at 6-60 units of modified activity of CX / liter of wash solution, preferably at

-50 unités d'activité régulière de Cx/litre de solu-  -50 units of regular activity of Cx / liter of solu-

tion de lavage ou à 12-30 unités d'activité modifiée  washing or 12-30 units of modified activity

de Cx/litre de solution de lavage.of Cx / liter of washing solution.

Selon une forme de réalisation préférée de l'invention, le pH de la solution de lavage se situe entre 7 et 10, de préférence, entre 7,5 et 9, 5, de telle sorte que la cellulase puisse exercer pleinement  According to a preferred embodiment of the invention, the pH of the wash solution is between 7 and 10, preferably between 7.5 and 9.5, so that the cellulase can fully exercise.

son activité.his activity.

Selon une forme de réalisation préférée du  According to a preferred embodiment of

procédé de lavage principal suivant la présente inven-  main washing process according to the present invention.

tion, on effectue une partie importante du lavage  tion, a large part of the washing is carried out

principal à une température inférieure à 700 C, de pré-  at a temperature below 700 C, preferably

férence, à une température inférieure à 600C.  at a temperature below 600C.

Les exemples ci-après illustrent davantage la fermentation au moyen de laquelle on obtient l'agent réducteur de dureté suivant l'invention (exemples 1, 6 et 8), de même que les compositions détergentes de lavage principal et les procédés de  The following examples further illustrate the fermentation by means of which the hardness reducing agent according to the invention (Examples 1, 6 and 8) is obtained, as well as the main wash detergent compositions and

lavage principal suivant la présente invention (exem-  main wash according to the present invention (eg

ples 2-8).ples 2-8).

Exemple 1Example 1

On cultive la souche DSM 1800 à 37 C sur un substrat dlagar-agar avec la composition suivante: Extrait de levure Difco 4 g K2HPO4 1 g MgSO4.7H20 0, 5 g Glucose 15 g Eau distillée 1.000 g  The strain DSM 1800 is cultured at 37 ° C. on a agar-agar substrate with the following composition: Difco yeast extract 4 g K 2 HPO 4 1 g MgSO 4 .7H 2 O 0.5 g Glucose 15 g Distilled water 1.000 g

Agar-agar 15 g.Agar-agar 15 g.

On prépare un premier groupe de ballons d'agitation de 500 ml contenant 100 ml d'un substrat de la composition suivante: Liqueur de macération de mais 2,4% Glucose 2,4% CaCO3 0,5% Huile de soya 0,5% Avant l'addition de CaC03, on règle le pH à 5,5 avec du NaOH 4N et on effectue la stérilisation à 121 C  A first group of 500 ml stirring flasks containing 100 ml of a substrate of the following composition was prepared: Corn maceration liquor 2.4% Glucose 2.4% CaCO3 0.5% Soybean oil 0.5 Before addition of CaCO 3, the pH is adjusted to 5.5 with 4N NaOH and the sterilization is carried out at 121 ° C.

pendant 20 minutes.for 20 minutes.

Après croissance pendant 7 jours sur une cultu-  After growing for 7 days on a cultural

re inclinée d'agar-agar, on transfère les spores dans  When agar-agar is inclined, the spores are transferred to

les ballons d'agitation à 500 ml du premier groupe.  the 500 ml stirring flasks of the first group.

On prépare un deuxième groupe de ballons d'agitation à 500 ml; chacun de ces ballons contient ml d'un substrat de la composition suivante:  A second group of 500 ml stirring flasks is prepared; each of these balloons contains ml of a substrate of the following composition:

NH4NO3 0,25%NH4NO3 0.25%

KH2PO4 0,56%KH2PO4 0.56%

K2HPO4 0,44%K2HPO4 0.44%

MgSO4.7H20 0,075% Cellulose -2,0% CaCO3 0,25% "Pluronic" 0,01% Avant l'addition de CaCO3, on règle le pH à 6,8-7 au moyen de NaOH 4N et on effectue la stérilisation par  MgSO4.7H20 0.075% Cellulose -2.0% CaCO3 0.25% "Pluronic" 0.01% Before the addition of CaCO3, the pH is adjusted to 6.8-7 using 4N NaOH and the sterilization is carried out by

traitement en autoclave à 1210C pendant ú0 minutes.  autoclave treatment at 12 ° C. for 100 minutes.

Après croissance pendant 2 fois 24 heures à 37 C dans le premier groupe de ballons d'agitation, on effectue l'inoculation avec 6% de ce premier groupe  After growth for 2 times 24 hours at 37 ° C. in the first group of stirring flasks, the inoculation is carried out with 6% of this first group.

de ballons dans le deuxième groupe.  balloons in the second group.

Après fermentation dans le deuxième groupe de ballons d'agitation-pendant 140 heures à 37 C, l'activité de cellulase est de 4,2 unités d'activitérégulière de Cx/ml de bouillon de culture. Ensuite,  After fermentation in the second group of stirring flasks for 140 hours at 37 ° C, the cellulase activity was 4.2 units of Cx / ml regular activity of culture broth. Then,

on purifie le bouillon de culture de la manière sui-  the broth is purified in the following manner

vante: on filtre le bouillon sur de la terre d'infu-  broth: we filter the broth on earth of infu-

soires ("Hyflo-supercell"), on concentre le filtrat  ("Hyflo-supercell"), concentrate the filtrate

par ultrafiltration et on lyophilise le produit rete-  by ultrafiltration and freeze-dried the product

nu sur le filtre. La poudre lyophilisée (A) a une activité de Cx de 745 unités d'activité régulière de x  naked on the filter. The lyophilized powder (A) has a Cx activity of 745 units of regular activity of x

Cx/g.Cx / g.

On effectue deux expériences exactement de la même manière que celle décrite ci-dessus. Après  Two experiments are performed in exactly the same manner as described above. After

fermentation dans le deuxième groupe de ballons dlagi-  fermentation in the second group of balloons

tation pendant 140 heures, l'activité régulière de cellulase C lors de ces deux expériences est de 2,2  during 140 hours, the regular activity of cellulase C in these two experiments is 2.2

et 6,0 unités de Cx/ml de bouillon de culture respec-  and 6.0 units of Cx / ml of culture broth resp.

tivement. De même, lors de ces deux expériences, la poudre lyophilisée a une activité régulière de Cx de  tively. Similarly, during these two experiments, the freeze-dried powder has a regular Cx activity of

558 unités (poudre B) et de 518 unités/g (poudre C).  558 units (powder B) and 518 units / g (powder C).

Exemple 2Example 2

Dans cet exemple, on utilise la préparation de cellulase en poudre A de l'exemple 1 comme agent  In this example, the cellulase powder preparation A of Example 1 is used as the

réducteur de dureté.hardness reducer.

On effectue l'expérience de lavage en procé-  The washing experiment is carried out by

dant à un lavage en une seule étape avec une machine à la-  washing in a single step with a washing machine.

ver à tambour entièrement automatique "IMIELE" conformément au programme de lavage appelé "Kogevask 95 C" indiqué dans la brochure "T-52001" décrivant la machine à laver "MIELE", type 421 S. "MIELE" est une marque commerciale. Le programme appelé "Kogevask 95 C"I a à peu près le profil température/durée ci-après: Durée (minutes) O 6 12 18 24 30 36 42 45  fully automatic drum worm "IMIELE" according to the washing program called "Kogevask 95 C" indicated in the brochure "T-52001" describing the washing machine "MIELE", type 421 S. "MIELE" is a trademark. The program called "Kogevask 95 C" I has approximately the following temperature / duration profile: Duration (minutes) O 6 12 18 24 30 36 42 45

Tempéra-tempera-

ture ( C) 16 27 43 61 76 85 85 82 rinça-  ture (C) 16 27 43 61 76 85 85 82 rinses

ge à l'eau froidein cold water

La solution de lavage a la composition sui-  The washing solution has the composition

vante: environ 16 litres d'eau d'une dureté d'envi-  about 16 liters of water with a hardness of about

ron 20 (selon les normes allemandes) contenant 5 g/ litre d'un détergent classique en poudre disponible  20 (according to German standards) containing 5 g / liter of a conventional powder detergent available

dans le commerce comportant les ingrédients princi-  in commerce containing the main ingredients

paux suivants (composition approximative en solides secs, poids/poids):  following levels (approximate composition in dry solids, weight / weight):

21% d'agents tensio-actifs (7% d'un alkyl-aryl-sulfo-  21% of surfactants (7% of an alkyl-aryl sulfoxide)

nate linéaire, 11% de savon et 3% de nonylphényl-  linear soap, 11% soap and 3% nonylphenyl

éthoxylate) 39% de charges (32% de tripolyphosphate de sodium, 4% de silicate de sodium et 3% de carbonate de sodium) 26% de perborate de sodium  ethoxylate) 39% fillers (32% sodium tripolyphosphate, 4% sodium silicate and 3% sodium carbonate) 26% sodium perborate

13% de sulfate de sodium.13% sodium sulfate.

A cette solution de lavage, on ajoute la pré-  To this washing solution is added the

paration enzymatique protéolytique-d"l'ALCALASE" en une concentration correspondant à 0,06 unité Anson/litre de solution. On détermine l'activité protéolytique de la préparation enzymatique conformément au procédé modifié d'Anson décrit dans "'Novo Analytical Method"  proteolytic enzymatic parenteration of ALCALASE in a concentration corresponding to 0.06 Anson units / liter of solution. The proteolytic activity of the enzyme preparation is determined according to the modified method of Anson described in "Novo Analytical Method"

N AF 4.3/5-GB (le procédé original d'Anson est dé-  N AF 4.3 / 5-GB (the original process of Anson is de-

crit dans "J. Gen. Physiol.", 22, 79-89 (1939)).  written in "J. Gen. Physiol.", 22, 79-89 (1939)).

A cette soluition de lavage, ou bien on n'ajoute rien ou on ajoute la préparation de cellulase de l'exemple 1 en une concentration correspondant à peu près à 40 unités d'activité régulière de Cx/litre  To this wash solution, either nothing is added or the cellulase preparation of Example 1 is added in a concentration corresponding to about 40 units of regular Cx / liter activity.

de solution.of solution.

Les matières d'essai utilisées pour chacune de ces deux expériences effectuées en parallèle sont les suivantes 1) 12 pièces d'un tissu-éponge blanc en coton mesurant 42 cm x 60 cm, et 2) 12 pièces de tissu interlock de coton blanc ayant les mêmes dimensions. De plus, avec les deux types de matières  The test materials used for each of these two experiments performed in parallel are as follows: 1) 12 pieces of a white cotton terry cloth measuring 42 cm x 60 cm, and 2) 12 pieces of white cotton interlock fabric having the same dimensions. In addition, with both types of materials

d'essai, on charge du linge propre en coton pour ob-  test, clean cotton linen is loaded to

tenir une lessive totale de 3 kg. Avant l'utilisa-  keep a total lye of 3 kg. Before using

tion, on lave 20 fois les matières d'essai avec un  tion, the test materials are washed 20 times with a

détergent classique contenant un perborate et en adop-  conventional detergent containing a perborate and

tant un programme à hautes températures.  both a high temperature program.

Après ces 20 lavages répétés qui simulent un lavage pratique des matières d'essai, ces dernières sont prêtes à subir l'expérience de lavage avec ou  After these 20 repeated washes which simulate a practical washing of the test materials, the latter are ready to undergo the washing experiment with or

sans cellulase.without cellulase.

On lave 6 fois les matières d'essai en pro-  The test materials are washed 6 times

cédant à des séchages intermédiaires sur une corde à linge pendant une nuit. Après le lavage final, on conditionne tous les échantillons d'essai (12 x 2 x 3)  yielding to intermediate drying on a clothesline for one night. After the final wash, all the test samples (12 x 2 x 3) are conditioned.

sur une corde à linge dans le même local.  on a clothesline in the same room.

L'évaluation est effectuée par un jury de membres. Sans connaître l'identité des différents échantillons d'essai, il a été demandé à chacune des dix premières personnes du jury de procéder à six évaluations (c'est-à-dire trois pour le tissu-éponge blanc et trois pour l'interlock de coton blanc) et, sur la base de ces évaluations, il a été possible de  The evaluation is done by a jury of members. Without knowing the identity of the different test samples, each of the first ten panelists was asked to make six evaluations (ie three for the white terry cloth and three for the interlock white cotton) and, on the basis of these assessments, it was possible to

sélectionner le jugement correct parmi ceux mention-  select the correct judgment from among those

nés ci-après: 1) le tissu traité avec la cellulase est  born as follows: 1) the cellulase-treated tissue is

plus doux que le tissu traité sans cellu-  softer than the fabric treated without

lase, 2) le tissu traité avec la cellulase a le même degré de douceur que le tissu'traité sans cellulase, et  lase, 2) cellulase-treated tissue has the same degree of softness as tissue without cellulase, and

248 1712248 1712

3) le tissu traité avec la cellulase est plus  3) the tissue treated with cellulase is no longer

dur que le tissu traité sans cellulase.  hard as the treated tissue without cellulase.

Parmi les 12 échantillons de tissu-épdnge de  Of the 12 samples of tissue-sponge of

coton traités à la cellulase, on a choisi trois échan-  treated with cellulase, three samples were selected

tillons de matière d'essai et, parmi les 12 échantil-  samples of test material and, among the 12 samples

lons de tissu-éponge blanc de coton traités sans cel-  lons of white cotton terry cloth treated without

lulase, on a également choisi trois échantillons de matière d'essai. On a disposé ces six échantillons en trois paires et l'on a demandé à chacun des dix premiers membres du jury d'évaluer séparément toutes les trois paires. On a procédé à des évaluations semblables avec l'interlock de coton blanc. Ensuite,  lulase, three samples of test material were also selected. These six samples were arranged in three pairs and each of the first ten members of the jury was asked to evaluate all three pairs separately. Similar assessments were made with the white cotton interlock. Then,

on a demandé aux dix autres personnes du jury d'éva-  the other ten people on the jury were asked to

luer le reste des matières d'essai de la même manière.  read the rest of the test materials in the same way.

En ce qui concerne uniquement le tissu-éponge  With regard only to the terry cloth

blanc, chaque membre du jury a procédé à trois évalua-  each member of the jury carried out three

tions, soit, pour tout le jury, un total de 60 évalua-  or, for the entire jury, a total of 60

tions pour cette matière d'essai.for this test substance.

L'interlock de coton blanc a été évalué de la même manière, le nombre total d'évaluations étant  The white cotton interlock was evaluated in the same way, the total number of assessments being

également, dans ce cas, de 60.also, in this case, 60.

Les résultats des évaluations décrites ci-  The results of the evaluations described below

dessus sont repris dans le tableau 1 ci-après. Ce tableau 1 indique également le pH de la solution détergente au début du lavage, après 10 minutes et  above are listed in Table 1 below. This table 1 also indicates the pH of the detergent solution at the beginning of the washing, after 10 minutes and

après 20 minutes (t = O, t = 10 et t = 20 respective-  after 20 minutes (t = 0, t = 10 and t = 20 respectively

ment).is lying).

Exemple 3Example 3

On a répété l'exemple 2. Les résultats obte-  Example 2 was repeated. The results obtained

nus sont repris dans le tableau 1.are shown in Table 1.

Exemple 4Example 4

Dans cet exemple, comme agent réducteur de dureté, on a utilisé la préparation de cellulase en poudre B de l'exemple 1. On a répété l'exemple 2, mais en y apportant les modifications suivantes v on a rempli la machine à laver avec la matière d'essai  In this example, as the hardness reducing agent, the powdered cellulase preparation B of Example 1 was used. Example 2 was repeated, but with the following modifications: the washing machine was filled with the test material

248 1712248 1712

comme décrit à l'exemple 2, puis on a chargé du linge en coton souillé pour obtenir, au total, une lessive  as described in Example 2, and then soiled cotton linen was loaded to obtain, in total, a laundry

de 3 kg. Le linge sale était constitué de sous-vête-  of 3 kg. The dirty laundry consisted of underclothes

ments en coton provenant d'un camp militaire. Les résultats sont repris dans le tableau 1.  cotton from a military camp. The results are shown in Table 1.

Exemple 5Example 5

Dans cet exemple, comme agent réducteur de dureté, on a utilisé la préparation de cellulase C de l'exemple 1. On a répété l'expérience décrite à l'exemple 2, mais en y apportant les modifications  In this example, as a hardness reducing agent, the cellulase preparation C of Example 1 was used. The experiment described in Example 2 was repeated, but with modifications.

suivantes: on a adopté le programme de lavage appe-  following: we adopted the washing program called

lé "Finvask 600ChI dans la brochure 'T-52001". Ce pro-  "Finvask 600ChI in the brochure 'T-52001'. This pro-

gramme présente le profil approximatif de température/ durée ci-après Durée (minutes) 0 6 12 18 24 30 36 42  gram has the approximate temperature / duration profile below Duration (minutes) 0 6 12 18 24 30 36 42

Tempéra-tempera-

ture (oC) 16 24 35 47 57 55 54 rinçage à l'eau froide La solution de lavage a un volume d'environ 26 litres. On règle légèrement le pH de la solution de lavage avec de l'acide citrique. Les résultats  ture (oC) 16 24 35 47 57 55 54 rinsing with cold water The washing solution has a volume of approximately 26 liters. The pH of the wash solution is slightly adjusted with citric acid. The results

obtenus sont repris dans le tableau 1.  obtained are shown in Table 1.

248 1712248 1712

TABLEAU 1TABLE 1

Exemple Matière Evaluation d'essai 73%: plus doux avec la cellulase Exemple 2 Tissu-éponge 2%: pas de différence pH à 25%: plus dur avec la t = 0: 10,0 cellulase t = 10 9,8 t = 20 9,7 Interlock 77%: plus doux avec la de coton cellulase 8%: pas de différence %: plus dur avec la _ cellulas e 68%: plus doux avec la cellulas e Exemple 3 Tissu-éponge 5%: pas de différence pH à 27%: plus dur avec la t = O 9,9. ..... cellulase t = 10 9,8 78%: plus doux avec la t = 20 9,6 Interlock cellulase de coton 2%: pas de différence %: plus dur avec la cellulase 84%: plus doux avec la cellulase Exemple 4 Tissu-éponge 3%: pas de différence pH à 13%: plus dur avec la t = 9,8 cellulase t = 10 9,7 82%: plus doux avec la t = 20 9, 6 cellulase Interlock 5%: pas de différence de coton 13%: plus dur avec la cellulas e  Example Material Test evaluation 73%: softer with cellulase Example 2 Terry cloth 2%: no difference pH at 25%: harder with t = 0: 10.0 cellulase t = 10 9.8 t = 20 9.7 Interlock 77%: softer with cellulase cotton 8%: no difference%: harder with cellulase e 68%: softer with cellulase e Example 3 Terry cloth 5%: no difference pH at 27%: harder with t = 0 9.9. ..... cellulase t = 10 9.8 78%: softer with t = 20 9.6 Cotton interlock cellulase 2%: no difference%: harder with cellulase 84%: softer with cellulase EXAMPLE 4 Sponge 3%: no difference pH 13%: harder with t = 9.8 cellulase t = 10 9.7 82%: softer with t = 20 9.6 Interlock cellulase 5%: no difference in cotton 13%: harder with cellulas e

248 1712248 1712

TABLEAU 1 (suite)TABLE 1 (continued)

Exemple 6Example 6

On cultive la souche DSM 1800 à 37 C sur un substrat d1agar-agar ayant la même composition que  The strain DSM 1800 is cultured at 37 ° C. on agar-agar substrate having the same composition as

celui de l'exemple -1.that of example -1.

On stérilise, à 121 C pendant 45 minutes, 300  At 121 ° C. for 45 minutes, 300

litres d'un substrat d'inoculation ou de préfermenta-  liters of an inoculation or pre-fermentation

tion de la composition suivante: Liqueur de macération de mais 2,4% Glucose 2,4% CaCO3 0,5% Huile de soya 0,4% (dont le pH avant l'addition de CaCO3 a été réglé à 5,5  Composition of the following composition: Corn maceration liquor 2.4% Glucose 2.4% CaCO3 0.5% Soybean oil 0.4% (whose pH before the addition of CaCO3 was adjusted to 5.5

à l'aide de NaOH 4N).with 4N NaOH).

Après croissance pendant 14 jours sur le subs-  After 14 days of growth on the

trat d'agar-agar, on transfère les spores dans les 300  agar-agar, the spores are transferred to the 300

litres du substrat stérilisé de préfermentation.  liters of the sterilized pre-fermentation substrate.

On fait passer de l'air stérile à travers le bouillon de préfermentation ainsi préparé pendant 49 heures à 37 C sous une pression de 0,5 atmosphère et à  Sterile air is passed through the pre-fermentation broth thus prepared for 49 hours at 37 ° C. under a pressure of 0.5 atmosphere and at room temperature.

raison de 300 litres/minute.300 liters / minute.

Par ébullition à 123 C pendant 60 minutes, on stérilise 350 litres d'un substrat de fermentation Exemple Matière Evaluation d'essai 93%: plus doux avec la cellulase Exemple 5 Tissu-éponge 0%: pas de différence pH à 7%: plus dur avec la t = O: 8,9 cellulase t = 10: 8,8 93%: plus doux avec la t = 30: 8,9 cellulase Interlock 2%: pas de différence de coton 3%: plus dur avec la cellulase principale de la composition suivante: Liqueur de macération de mais 10% Cellulose 3% CaCO3 0,25%  By boiling at 123 ° C. for 60 minutes, 350 liters of a fermentation substrate are sterilized. Example Material Test evaluation 93%: softer with cellulase Example 5 Terry cloth 0%: no difference pH at 7%: more hard with t = O: 8.9 cellulase t = 10: 8.8 93%: softer with t = 30: 8.9 cellulase Interlock 2%: no difference in cotton 3%: harder with cellulase composition of the following composition: Maize maceration liquor 10% Cellulose 3% CaCO3 0.25%

NH4NO3 0,25%NH4NO3 0.25%

KH2PO4 0,56%KH2PO4 0.56%

K 2HPO4 0,44%K 2HPO4 0.44%

2 4 v4 MgS04 0,08% "Pluronic"t 0,014% (dont le pH avant l'addition de CaC03 a été réglé à 6,8-7  2 4 v4 MgSO 4 0.08% "Pluronic" t 0.014% (whose pH before the addition of CaCO 3 was adjusted to 6.8-7

à l'aide de NaOH 4N).with 4N NaOH).

On transfère 35 litres du bouillon de préfermen-  35 liters of pre-fermentation broth are transferred

tation dans le substrat ainsi stérilisé de fermentation principale. On fait passer de l'air stérile à travers le bouillon ainsi préparé de fermentation principale sous une pression de 0,5 atmosphère et à raison de 300 1/minute. On maintient le pH sous 7 avec addition de KH2PO4. On effectue la fermentation à 37 C pendant 136 heures. Au terme de la fermentation, l'activité est  in the sterilized substrate of the main fermentation. Sterile air is passed through the broth thus prepared for main fermentation under a pressure of 0.5 atmosphere and at a rate of 300 l / minute. The pH is maintained at 7 with the addition of KH2PO4. The fermentation is carried out at 37 ° C. for 136 hours. At the end of the fermentation, the activity is

de 29 unités d'activité régulière de Cx/ml.  of 29 units of regular activity of Cx / ml.

On purifie le bouillon de culture de la manière suivante: On filtre le bouillon de culture sur de la terre  The culture broth is purified in the following manner: The culture broth is filtered on soil

d'infusoires ("Hyflo-supercell"). On concentre le fil-  infusoria ("Hyflo-supercell"). The focus is

trat par ultrafiltration et on lyophilise le produit retenu sur le filtre (qui, pour plus de brièveté, sera  treated by ultrafiltration and freeze-dried the product retained on the filter (which, for brevity, will be

appelé ci-après "produit retenu D").  hereinafter referred to as "retained product D").

La poudre lyophilisée D a une activité régu-  The lyophilized powder D has a regular activity

lière de Cx de 1.316 unités/g et une activité modifiée  of 1.316 units / g and a modified activity

de Cx de 953 unités/g.Cx of 953 units / g.

De la même manière que celle indiquée ci-  In the same way as indicated above

dessus, on effectue deux fermentations supplémentai-  above, two additional fermentations are carried out

res avec des ultrafiltrations ultérieures, Les pro-  with subsequent ultrafiltrations,

duits retenus correspondants sont appelés "produits retenus E et F". On lyophilise le produit retenu F et la poudre lyophilisée F a une activité régulière de Cx de 743 unités/g et une activité modifiée de Cx  The corresponding retained products are called "selected products E and F". The retained product F is lyophilized and the freeze-dried powder F has a regular Cx activity of 743 units / g and a modified Cx activity.

248 1712248 1712

de 580 unités/g.of 580 units / g.

On sépare la poudre lyophilisée D en compo-  The freeze-dried powder D is separated into

sants différents par chromatographie dl'échange d'a-  different by chromatography of the exchange of

nions de la manière suivante: on en dissout 100 g dans 2.000 ml dleau distillée à 4 C. On règle le pH à 7,5 avec le tampon "tris" 1M. Ensuite, on applique la solution dans une colonne échangeuse d'ions de  In the following manner, 100 g are dissolved in 2,000 ml of distilled water at 4 ° C. The pH is adjusted to 7.5 with the 1M tris buffer. Then the solution is applied in an ion exchange column of

4.000 ml avec 100 g de "DEAE-Sephadex A-50". La fi-  4.000 ml with 100 g of "DEAE-Sephadex A-50". The

gure 1 montre la capacité d'absorption OD280 à 280 nm des fractions libérées de la colonne. On élue les protéines fixées dans la colonne par un gradient de NaCl; on mesure la concentration de NaCl et on en trace une courbe dans le diagramme de la figure 1  Figure 1 shows OD280 absorption capacity at 280 nm of the fractions released from the column. Proteins fixed in the column are eluted by a NaCl gradient; the concentration of NaCl is measured and a curve is plotted in the diagram of FIG.

conjointement avec la capacité d'absorption des frac-  together with the absorption capacity of the fractions

tions.tions.

On forme des groupes des fractions contenant des protéines comme indiqué par les crochets de la figure 1 et les fractions groupées sont numérotées de 1 à 5. Ensuite, on lyophilise séparément les groupes  Fractions containing proteins are formed as indicated by the square brackets of Figure 1 and the pooled fractions are numbered from 1 to 5. The groups are then lyophilized separately.

de fractions 1 à 5.fractions 1 to 5.

Les groupes de fractions lyophilisées sont  The lyophilized fraction groups are

caractérisés par la quantité de protéines, la compo-  characterized by the amount of protein, the composition

sition des protéines et l'activité régulière de Cx (voir tableau 2 ciaprès). On détermine le point isoélectrique des protéines avec un appareil "LKB multiphoer".  protein and regular Cx activity (see Table 2 below). The isoelectric point of the proteins is determined with a "LKB multiphoer" apparatus.

TABLEAU 2TABLE 2

Protéines Nombre Inter- Ren- Rende-  Protein Number Inter- Ren-

(g) total valle dement ment d'uni- du pH en en C x tés protéi- (%) d'actines vité ré- (%) gulière de C Matière de départ 53,0 131.600 3,5-9,0 Groupe 1*) 8,6 47.400 6-9,0 16 36 Groupe 2 0,9 1.000 4-6 2 1 Groupe 3 4,8 9.500 4-5,5 9 7 Groupe 4 7,1 11.000 3,5-5,5 13 8 Groupe 5 17,7 18.600 3,55 33 14 [I______ _Total 73 66  (g) total amount of pH from one to two (C)% protein content (%) of reactants C (R) content (%) starting material 53.0 131.600 3.5-9.0 Group 1 *) 8.6 47.400 6-9.0 16 36 Group 2 0.9 1,000 4-6 2 1 Group 3 4.8 9.500 4-5.5 9 7 Group 4 7.1 11.000 3.5-5, 5 13 8 Group 5 17.7 18.600 3.55 33 14 [I_______Total 73 66

*) (= fraction ACxI).*) (= ACxI fraction).

Le groupe 1 contient tous les calins, c'est-à-dire les protéines qui fixées à la résine échangeuse d'anions  Group 1 contains all hugs, that is, the proteins that attach to the anion exchange resin

composants al-components

ne sont pasare not

à 6,5 t pH t 7,5.at 6.5 t pH t 7.5.

Dès lors, la quantité totale d'activité de cellulase modifiée C est présente dans le groupe 1. qui contient également toutes les activités cellulolytiques d'un  Therefore, the total amount of modified cellulase activity C is present in group 1. which also contains all the cellulolytic activities of a

pH supérieur à 6.pH greater than 6.

Afin d'évaluer lequel des cinq groupes con-  In order to evaluate which of the five groups

tient la fraction exerçant l'effet adoucissant le plus  holds the fraction exerting the most softening effect

intense, on effectue les expériences décrites ci-  intense, the experiments described above are carried out

après.after.

On soumet un tissu-éponge de coton à 20 pré-  A 20-year-old cotton terry cloth is

lavages comme décrit à l'exemple 2. Après ces pré-  washes as described in Example 2. After these

lavages, on découpe des échantillons de 10 cm x 10 cm  washes, cut 10 cm x 10 cm samples

(environ 4 g) et on les marque.(about 4 g) and mark them.

On-effectue ensuite le traitement à la cel-  The treatment is then carried out

lulase dans une machine à laver de laboratoire "Terg-O-Tometer" dans les conditions suivantes: Détergent et dosage: comme indiqué à l'exemple 2, à l'exception du dosage de la cellulase pH initial: Température: Durée: Dureté de l'eau: Volume par becher: Nombre d'échantillons par becher: Dosage de la cellulase: environ 9,5 500C minutes  lulase in a laboratory washing machine "Terg-O-Tometer" under the following conditions: Detergent and dosage: as indicated in Example 2, with the exception of the cellulase assay initial pH: Temperature: Duration: Hardness of water: Volume per beaker: Number of samples per beaker: Determination of cellulase: about 9.5 500C minutes

(normes al-  (standards

lemandes) 1.200 mlthe requests) 1.200 ml

voir tableauSee table

ci-dessous; dans tous les cas:below; in all cases:

mg des grou-mg of groups

pes lyophilisés/ litre Dosage: Groupe Groupe Groupe Groupe Groupe mg de protéine/l 1 2 3r 4 5  lyophilized pes / liter Dosage: Group Group Group Group Group mg of protein / l 1 2 3r 4 5

1 2 3 4 $1 2 3 4 $

31 13 28 34 5231 13 28 34 52

Unités d'activité régulière de Cx/l 170 15 56 52 55 Unités d'activité modifiée de Cx/1 170 O O O j O L'évaluation est effectuée par un jury de  Regular activity units of Cx / l 170 15 56 52 55 Modified activity units of Cx / 1 170 O O O j O The evaluation is carried out by a jury of

10 membres auxquels on a donné différents jeux d'échan-  10 members who were given different games of exchange.

tillons constitués chacun de 3 échantillons et on leur a demandé de classer les échantillons de chaque groupe  samples each consisting of 3 samples and asked to rank the samples from each group

en fonction de leur douceur. Chaque jeu était cons-  according to their softness. Each game was

titué de deux échantillons que l'on avait lavés avec un des groupes, ainsi que d'un échantillon faisant office de témoin. Sur les dix membres du jury, neuf I I I I i ont trouvé que les échantillons lavés avec le groupe 1 étaient plus doux que celui auquel on n'avait pas ajouté le groupe. Tous les autres groupes n'ont pas  of two samples that had been washed with one of the groups, as well as a sample serving as a control. Of the ten members of the jury, nine found that the samples washed with Group 1 were milder than the one to which the group had not been added. All other groups do not have

adouci sensiblement les échantillons.  softened the samples substantially.

D'après l'exposé ci-dessus, on peut conclure que le groupe 1 contient l'activité de C exerçant x un effet adoucissant. L'activité de Cx des groupes 3-5 n'exerce aucun effet adoucissant important à des  From the above discussion, it can be concluded that group 1 contains the activity of C exerting a softening effect. The Cx activity of groups 3-5 has no significant softening effect on

dosages auxquels cet effet s'exerce normalement.  dosages to which this effect is normally applied.

Exemple 7Example 7

On soumet le produit retenu E de l'exemple 6  The retained product E of Example 6 is subjected

à un séchage par pulvérisation afin de former un con-  spray drying to form a

centrat brut.gross center.

On fait passer ce concentrat brut à travers un tamis dont les trous ont un diamètre de 0,5- mm,  This crude concentrate is passed through a sieve whose holes have a diameter of 0.5 mm,

pour obtenir ainsi un diamètre moyen de 17,4 r.  to obtain an average diameter of 17.4 r.

L'activité de cellulase du concentrat dans cet état  The cellulase activity of the concentrate in this state

est de 650 unités d'activité régulière de Cx/g.  is 650 units of regular activity of Cx / g.

Dans un mélangeur ("L6dige FM 50/1MZ") tour-  In a mixer ("L6dige FM 50 / 1MZ") turn

nant à une vitesse de 120 tours/minute et dont la lame  at a speed of 120 rpm and whose blade

* tourne à 3.000 tours/minute, on malaxe un mélange com-* rotates at 3,000 rpm, kneading a mixture of

prenant: le concentrat de cellulase ci-dessus 2,5 kg Poudre de cellulose ("Arbocel BC 200") 1,0 kg TiO2 0,2 kg  taking: cellulase concentrate above 2.5 kg Cellulose powder ("Arbocel BC 200") 1.0 kg TiO2 0.2 kg

NaCl (granularité des particules: <0,2 mm) 6,1 kg.  NaCl (granularity of the particles: <0.2 mm) 6.1 kg.

Au mélange ainsi obtenu, on ajoute 1 kg d'une solution aqueuse à 20% de polyvinylpyrrolidone (PVP K 30) de façon à maintenir les vitesses de rotation indiquées ci-dessus pour le mélangeur et la lame. On effectue  To the mixture thus obtained, 1 kg of a 20% aqueous solution of polyvinylpyrrolidone (PVP K 30) was added so as to maintain the rotational speeds indicated above for the mixer and the blade. We perform

l'addition de la solution aqueuse au moyen d'un aju-  addition of the aqueous solution by means of an addition

tage à pression et cette addition exige une durée d'une minute. Ensuite, on soumet ce mélange humide à un traitement complémentaire pendant 3 minutes dans le mélangeur tournant à 200 tours/minute et dont la lame tourne à 3.000 tours/minute. On ajoute deux fois  pressure and this addition requires a period of one minute. Then, this wet mixture is subjected to a complementary treatment for 3 minutes in the mixer rotating at 200 rpm and whose blade rotates at 3,000 rpm. We add twice

une quantité supplémentaire d'eau (à savoir respec-  additional quantity of water (ie

tivement 200 et 100 g d'eau) on procédant chaque fois à un traitement ultérieur d'une durée de 3 minutes dans le mélangeur. On observe que la température de la masse s'élève de 25 à 30 C au cours de la granula-  200 to 100 g of water), each subsequent treatment is carried out for 3 minutes in the mixer. It is observed that the temperature of the mass rises from 25 to 30 C during the granulation.

tion. On sèche les granulés humides dans un lit flui-  tion. The wet granules are dried in a fluid bed.

de jusqu'à une teneur en eau de 1,6%. Les expressions  from up to a water content of 1.6%. The expressions

"L6dige" et "Arbocel" désignent des marques commercia-  "L6dige" and "Arbocel" refer to trademarks

les. Activité régulière de C des granulés secs: Calculé 164 units d'activité régulière de C/g Trouvé 164 unités d'activité régulière de Cx/g Trouvé 136 unités d'activité régulière de Cl/g  the. Regular activity of C dry granules: Calculated 164 units of regular activity of C / g Found 164 units of regular activity of Cx / g Found 136 units of regular activity of Cl / g

Perte d'activité: 17%.Loss of activity: 17%.

Ce produit se présente sous forme de parti-  This product is in the form of

cules lenticulaires. Il sera appelé plus brièvement  lenticular cells. It will be called more briefly

"granulé E"."granulated E".

La granulométrie de ce granulé E est la sui-  The granulometry of this granulate E is as follows

vante: Granularité des particules (m) % de particules  vante: Granularity of particles (m)% of particles

>1.180 15> 1.180 15

>1.000 28> 1.000 28

> 850 43> 850 43

> 707 62> 707 62

> 600 76> 600 76

>500 87> 500 87

>420 94> 420 94

> 300 1,8> 300 1.8

Diamètre moyen = 790,m.Average diameter = 790, m.

On enrobe le granulé E de la manière décrite  The granule E is coated in the manner described

ci-après. On verse 4 kg du granulé E dans un mélan-  below. 4 kg of granule E are poured into a mixture of

geur ('L6dige"l, type M 20E). On chauffe le mélange à 650C, puis on ajoute 160 g de polyéthylène-glycol 4000 fondu (poids moléculaire: 4.000); on malaxe ce mélange pendant 1 minute. Ensuite, on saupoudre le  The mixture is heated to 650 ° C, then 160 g of molten polyethylene glycol 4000 (molecular weight: 4,000) are added, the mixture is kneaded for 1 minute, and the mixture is then dusted.

granulé avec 360 g de TiO2 et 90 g de silicate de ma-  granulated with 360 g of TiO2 and 90 g of magnesium silicate

gnésium. Après un temps de mélange de 5 minutes, on verse encore 40 g de polyéthylène-glycol 4000 sur le  sium. After a mixing time of 5 minutes, another 40 g of polyethylene glycol 4000 are poured onto the

mélange et l'on poursuit le malaxage pendant une minu-  mixing and mixing is continued for one minute.

te. On laisse refroidir la charge. Pendant 1,5 minu-  you. The charge is allowed to cool. For 1.5 minutes

te, à une température de 650C, on saupoudre les granu-  at a temperature of 650C, the granules are sprinkled

lés enrobés ainsi obtenus sur un lit fluide. L'analyse suivante fait ressortir l'activité  the coated thus obtained on a fluid bed. The following analysis highlights the activity

du granulé.granulate.

Le granulé enrobé, le granulé enrobé et sau-  The coated granulate, the coated granulate and sau-

poudré, de même que le granulé E indiqués ci-dessus sont des formes de réalisation spécifiquesd'un agent  powder, as well as the granules E indicated above are specific embodiments of an agent

réducteur de dureté sous forme d'un additif pour dé-  hardness reducer in the form of an additive for

tergent. On effectue une expérience de lavage avec le  detergent. A washing experiment is carried out with the

granulé E comme décrit à l'exemple 6, mais en appor-  granulate E as described in Example 6, but in

tant les modifications suivantes Dosage de la cellulase  both the following changes Cellulase dosage

333, 667, 1.000, 1.333 et 1.667 mg/litre.  333, 667, 1,000, 1,333 and 1,667 mg / liter.

On effectue l'évaluation comme décrit à l'e-  The evaluation is carried out as described in

xemple 6, mais avec les changements suivants: Chaque jeu d'échantillons est constitué de deux échantillons, à savoir un échantillon lavé avec  xample 6, but with the following changes: Each set of samples consists of two samples, namely a sample washed with

la solution détergente contenant de la cellulase ren-  the detergent solution containing cellulase

due inactive et un échantillon lavé avec la solution  inactive and a sample washed with the solution

détergente contenant de la cellulase active. On ob-  detergent containing active cellulase. We ob-

tient la cellulase rendue inactive en préparant une solution mère avec 10 g du granulé E dissous dans l'eau à 250 ml, puis en chauffant cette solution.à 901C et en maintenant cette température pendant 10-15 minutes. Au total, on évalue 5 jeux; les résultats des évaluations sont repris dans le tableau 3 ci-après: Granulé Granulé enrobé et enrobé saupoudré Unités d'activité régulière de Cx/g 137 138  Hold the cellulase inactivated by preparing a stock solution with 10 g of the E granules dissolved in water at 250 ml, and then heating this solution to 90 ° C. and maintaining this temperature for 10-15 minutes. In total, we evaluate 5 games; the results of the evaluations are given in Table 3 below: Granulated Pelleted and powder coated granules Regular activity units of Cx / g 137 138

TABLEAU 3TABLE 3

Evaluation de la douceur (%)Evaluation of softness (%)

Exemple 8Example 8

Dans cet exemple, on compare différentes sou-  In this example, we compare different

ches d'Humicola insolens à savoir les souches CBS 14764 (identique à la souche ATCC 22082), DSM 2069 et DSM 2068). On cultive ces souches comme indiqué à l'exemple 1, avec cette exception qu'en plus  Humicola insolens, namely strains CBS 14764 (identical to strain ATCC 22082), DSM 2069 and DSM 2068). These strains are cultured as indicated in Example 1, with the exception that in addition

des constituants indiqués dans cet exemple 1, le subs-  constituents indicated in this example 1, the subset

trat du deuxième groupe de ballons d'agitation con-  of the second group of stirring flasks con-

tient 14% d'alcool de mais. On forme les poudres lyo-  holds 14% of corn alcohol. Lyophilic powders are formed

philisées à partir des bouillons de fermentation comme  philized from fermentation broths like

indiqué à l'exemple 1. On élimine les sels des pou-  Example 1 is removed. The salts of the

dres lyophilisées, on les redissout et on en élimine à nouveau les sels dans une colonne de "Sephadex G 25",  Once freeze-dried, they are redissolved and the salts are removed again in a "Sephadex G 25" column.

puis on les lyophilise à nouveau. Ces poudres lyo-  then freeze-dried again. These lyophilic powders

philisées ont respectivement des activités régulières  philized have regular activities respectively

de Cx de 1.096 unités, 480 unités et 690 unités/g.  Cx of 1,096 units, 480 units and 690 units / g.

De même, dans cet exemple, on utilise une portion de  Similarly, in this example, we use a portion of

la poudre lyophilisée F de l'exemple 6 (DSM 1800).  lyophilized powder F of Example 6 (DSM 1800).

On soumet le groupe lyophilisé 1 de ltexem-  The freeze-dried group 1 of ltexem-

ple 6 à une purification complémentaire par échange de cations sur "CMSepharosc CL 6B". On dissout ce groupe lyophilisé 1 en une quantité correspondant à Jeu n0 Dosage de la Cellulase | Cellulase inactive  pl 6 to further purification by cation exchange on "CMSepharosc CL 6B". This freeze-dried group 1 is dissolved in an amount corresponding to FIG. Inactive cellulase

cellulase Evalution: l'échantillon trai-  cellulase Evalution: the sample treated

(mg/litre) té comme indiqué est le plus doux, à savoir:  (mg / liter) as indicated is the mildest, namely:

1 333 60% 40%1 333 60% 40%

2 667 70% 30%2,667 70% 30%

3 1.000 70% 30%3 1,000 70% 30%

4 1.333 70% 30%4 1.333 70% 30%

1.667 90% 10%1.667 90% 10%

1,7 g de protéine dans un tampon d'acide citrique à un pH de 5 et on applique la solution ainsi obtenue dans une colonne de 1.000 ml contenant lléchangeur de cations sur lequel s'est fixée la protéine. On élue avec un gradient de NaCl et-, à une concentration en NaCl de 250 mM, on élue la protéine ACxI active de carboxyméthyl-cellulose pure. Par électrophorèse sur gel de SDS, on constate que le poids moléculaire de cette protéine est de 80.000. On utilise également cette protéine pour la préparation dtantisérum (lapins). On utilise ensuite cet antisérum pour la quantification de la teneur de la protéine ACxI au moyen de techniques immunologiques, c'est-à-dire l'limmunoélectrophorèse en fusées, comme décrit par B. Weeke dans "Scandinavian I. Immun.1" 2 suppl. 1,  1.7 g of protein in a citric acid buffer at pH 5 and the solution thus obtained was applied in a 1000 ml column containing the cation exchanger to which the protein was attached. It is eluted with a NaCl gradient and at a concentration of 250 mM NaCl the active ACxI protein is eluted with pure carboxymethylcellulose. By SDS gel electrophoresis, it is found that the molecular weight of this protein is 80,000. This protein is also used for the preparation of antiserum (rabbits). This antiserum is then used for quantification of the ACxI protein content using immunological techniques, i.e., rocket immunoelectrophoresis, as described by B. Weeke in "Scandinavian I. Immun.1" 2 Supp. 1

page 37 (1973).page 37 (1973).

La figure 2 donne 1image de limmunoélectro-  Figure 2 shows the image of immunoelectron

phorèse en fusées. On effectue cette électrophorèse pendant une nuit dans un tampon de tris-maléate d'un  rocket phoresis. This electrophoresis is carried out overnight in a tris-maleate buffer of

pH de 7 et avec un gradient de tension de 1 volt/cm.  pH 7 and with a voltage gradient of 1 volt / cm.

En figure 2, a correspond au produit DSM 2069 (3 mg/ml), b correspond au produit CBS 14764 (2 mg/ml), c correspond au produit DSM 2068 (3 mg/ml) et dn correspond au produit DSM 1800  In FIG. 2, a corresponds to the product DSM 2069 (3 mg / ml), b corresponds to the product CBS 14764 (2 mg / ml), c corresponds to the product DSM 2068 (3 mg / ml) and dn corresponds to the product DSM 1800

(n mg/ml).(n mg / ml).

Les préparations de cellulase indiquées ci-  The cellulase preparations indicated below

dessus et provenant des produits DSM 1800, CBS 14764, DSM 2068 et DSM 2069 ont été utilisées dans l'expérience de lavage décrite ci-dessous:  above and from products DSM 1800, CBS 14764, DSM 2068 and DSM 2069 were used in the washing experiment described below:

On soumet un tissu-éponge de coton à 20 pré-  A 20-year-old cotton terry cloth is

lavages comme décrit à l'exemple 2. Après ces pré-  washes as described in Example 2. After these

lavages, on découpe des échantillons de 10 cm x 10 cm  washes, cut 10 cm x 10 cm samples

(environ 4 g) et on les marque.(about 4 g) and mark them.

On effectue ensuite le traitement à la cel-  The treatment is then carried out

lulase dans une machine à laver de laboratoire "ITerg-O-Tometer" dans les conditions suivantes: Détergent et dosage: comme indiqué à l'exemple 2, à l'exception du dosage de la cellulase pH initial: environ 9,5 Température: 50 C Durée: 30 minutes  lulase in a laboratory washing machine "ITerg-O-Tometer" under the following conditions: Detergent and dosage: as indicated in Example 2, with the exception of the cellulase dosage initial pH: about 9.5 Temperature: 50 C Duration: 30 minutes

Dureté de l'eau 20 (normes alle-Hardness of water 20 (German standards

mandes) Volume par becher: -1.200 ml Nombre d'échantillons par becher: 10 Dosage de la cellulase: voir tableau ci-après  per beaker: -1,200 ml Number of samples per beaker: 10 Determination of cellulase: see table below

On ajoute la préparation de cellulase prove-  The cellulase preparation

nant du produit DSM 1800 en un dosage de 76 unités d'activité régulière de Cx/litre et l'on ajoute les autres préparations de cellulase en un dosage donnant, dans la solution de lavage, la même concentration de protéine ACxI (déterminée par voie immunologique comme décrit ci-dessus dans le présent exemple) que dans la  from the DSM 1800 product in a dosage of 76 units of regular Cx / L activity and the other cellulase preparations are added in an assay giving the same concentration of ACxI protein in the wash solution (determined by immunologically as described above in the present example) that in the

préparation DSM 1800. On répète six fois les traite-  preparation DSM 1800. The treatment is repeated six times

ments à-la cellulase. Après chaque traitement, on lave convenablement les échantillons dans de l'eau du robinet et on les sèche à l'air pendant une nuit  cellulase. After each treatment, the samples are washed well in tap water and air-dried overnight.

sur une corde à linge.on a clothesline.

L'évaluation est effectuée par un jury de membres auxquels on a donné quatre jeux différents  The evaluation is done by a jury of members who have been given four different games

d'échantillons constitués chacun de trois échantil-  samples consisting of three samples each

lons et on leur a demandé de classer Ies échantil-  and asked them to rank the samples

Dosage de la Néant DSM CBS DSM DSM cellulase -1800 14764 2068 2069 (mg/litre) O 102 39 96 o105  DSM CBS DSM DSM cellulase -1800 Absent Assay 14764 2068 2069 (mg / liter) O 102 39 96 o105

Unités dt'acti-Units of activity

vité régulièreregular life

de Cx/litre 76 43 66 50 -of Cx / liter 76 43 66 50 -

_ _ _ _ _ _ _ _ ___ a___ à _ _ I__ _ _ _ _ _ _ _ ___ a___ to _ _ I_

lons de chaque jeu en fonction de leur douceur.  each game according to their softness.

A l'échantillon le plus doux d'un jeu, on a attribué le chiffre "1", à l'échantillon suivant, le chiffre 1"2" et ainsi de suite. Le tableau 4 ciaprès donne la composition des jeux évalués, de même que les ré-  To the sweetest sample of a game, the number "1" has been assigned to the next sample, the number 1 "2" and so on. Table 4 below gives the composition of the games evaluated, as well as the

sultats des évaluations, exprimés en pour-cent (pour-  results of the evaluations, expressed as a percentage (

centage du jury rangeant un certain échantillon dans une certaine catégorie de douceur):  percentage of the jury ranking a certain sample in a certain category of softness):

TABLEAU 4TABLE 4

Evaluation de la douceur (%) Jeu n Cellulase Catégorie de douceur  Evaluation of softness (%) Game n Cellulase Category of softness

échan-exchanged

tillon le plus doux 1 Néant 0 10 90 Poudre C (exemple 6) 80 20 0 Poudre de CBS 20 70 10 2 Néant 0 30 70 Poudre C (exemple 6) 70 30 0 Poudre de DSM 2068 30 40 30 3 Néant 0 20 80 Poudre C (exemple 6) 30 50 20 Poudre de DSM2069 70 30 O  Mildest sweep 1 None 0 10 90 Powder C (Example 6) 80 20 0 Powder CBS 20 70 10 2 None 0 30 70 Powder C (Example 6) 70 30 0 Powder DSM 2068 30 40 30 3 None 0 20 80 Powder C (Example 6) 30 50 20 Powder of DSM2069 70 30 O

....,..DTD:...., .. DTD:

Claims (18)

REVENDICATIONS 1. Agent réducteur de dureté sous forme d'un additif pour une composition détergente, un composant essentiel de cet agent étant une cellulase fongique, caractérisé en ce que la cellulase fongique peut être produite par une souche d'Humicola insolens (Humicola grisea var. thermoidea), tandis que l'agent réducteur de dureté est destiné à une composition  A hardness reducing agent in the form of an additive for a detergent composition, an essential component of this agent being a fungal cellulase, characterized in that the fungal cellulase can be produced by a strain of Humicola insolens (Humicola grisea var. thermoidea), while the hardness reducing agent is intended for a composition détergente pour un lavage principal.  detergent for a main wash. 2. Agent réducteur de dureté suivant la revendication 1, caractérisé en ce que la souche d'Humicola insolens est celle cld6posée sous.le n   2. hardness reducing agent according to claim 1, characterized in that the strain of Humicola insolens is that deposited under n DSM 1800.DSM 1800. 3. Agent réducteur de dureté suivant l'une  3. Hardness reducing agent according to one quelconque des revendications 1 et 2, caractérisé en  any of claims 1 and 2, characterized in that ce que la cellulase fongique est enrichie vis-à-vis de la fraction de cellulase qui ne se fixe pas à un  that fungal cellulase is enriched with respect to the fraction of cellulase that does not attach to a échangeur d'anions à 6,5 ' pH - 7,5.  anion exchanger at 6.5 'pH - 7.5. 4. Agent réducteur de dureté suivant la revendication 3, caractérisé en ce que la cellulase fongique présente un rapport (activité modifiée de  Hardness reducing agent according to claim 3, characterized in that the fungal cellulase has a ratio (modified activity of Cx/activité régulière de Cx) d'au moins 0,6, de pré-  Cx / regular activity of Cx) of at least 0.6, pre- férence, d'au moins 0,8.at least 0.8. 5. Agent réducteur de dureté suivant l'une  5. Hardness reducing agent according to one quelconque des revendications 1 à 4, caractérisé en  any of claims 1 to 4, characterized in ce qu'il est un granulé ne dégageant pas de poussiè-  it is a granulate that does not give off dust res. 6. Agent réducteur de dureté suivant la revendication 5, caractérisé en ce que le granulé ne dégageant pas de poussières est enrobé d'un agent de blanchiment en combinaison avec un agent fixant  res. 6. hardness reducing agent according to claim 5, characterized in that the non-dusting granule is coated with a bleaching agent in combination with a fixing agent. les poussières.dust. 7. Agent réducteur de dureté suivant la revendication 6, caractérisé en ce que l'agent de  7. hardness reducing agent according to claim 6, characterized in that the agent of blanchiment est le TiO2.bleaching is TiO2. Z 4 8 1 7 1 2Z 4 8 1 7 1 2 8. Agent réducteur de dureté suivant l'une  8. Hardness reducing agent according to one quelconque des revendications 1 à 4, caractérisé en  any of claims 1 to 4, characterized in ce qu'il est un liquide, tandis que la cellulase fon-  it's a liquid, whereas cellulase makes gique est prévue sous forme d'un concentrat de cel-  is planned in the form of a concentrate of this lulase en suspension dans un agent tensio-actif non ionique. 9. Agent réducteur de dureté suivant la revendication 8, caractérisé en ce qu'il contient  lulase suspended in a nonionic surfactant. 9. A hardness reducing agent according to claim 8, characterized in that it contains également un agent épaississant.also a thickening agent. 10. Agent réducteur de dureté suivant l'une  10. Hardness reducing agent according to one quelconque des revendications 1 à 4, caractérisé en  any of claims 1 to 4, characterized in ce qu'il est un liquide, tandis que la cellulase fon-  it's a liquid, whereas cellulase makes gique est prévue sous forme d'un concentrat de cellu-  is provided in the form of a cell concentrate. lase en solution dans un milieu aqueux, de préférence,  dissolved in an aqueous medium, preferably en présence d'un agent stabilisant.  in the presence of a stabilizing agent. Il. Composition détergente pour lavage prin-  He. Detergent composition for primary washing cipal, caractérisée en ce qu'elle comprend des ingré-  characterized in that it includes dients détergents et un agent réducteur de dureté  detergents and a hardness reducing agent suivant l'une quelconque des revendications l à 10 en  according to any one of claims 1 to 10 in une quantité réduisant la dureté de telle sorte que  a quantity reducing the hardness so that le pH d'une solution de 1 g de cette composition dé-  the pH of a solution of 1 g of this composition de- tergente de lavage principal dans un litre d'eau d'une dureté de 100 (selon les normes allemandes) avant l'addition de cette composition détergente de lavage principal, se situe dans l'intervalle allant de 7 à 10, de préférence, dans l'intervalle allant  The main washing solution in one liter of water with a hardness of 100 (according to German standards) before the addition of this main washing detergent composition, is in the range of 7 to 10, preferably in the interval going de 7,5 à 9,5.from 7.5 to 9.5. 12. Composition détergente de lavage prin-  12. Washing detergent composition primarily cipal suivant la revendication 11, caractérisée en ce que l'agent réducteur de dureté est présent en une quantité correspondant à 2,5-100 unités d'activité  cipal according to claim 11, characterized in that the hardness reducing agent is present in an amount corresponding to 2.5-100 activity units. régulière de Cx/g de la composition détergente de la-  of Cx / g of the detergent composition of the vage principal ou à 1,5-60 unités d'activité modifiée  or 1.5-60 units of modified activity de Cx/g de la composition détergente de lavage prin-  of Cx / g of the washing detergent composition cipal.cipal. aat 13. Composition détergente de lavage prin-  13. Main washing detergent composition cipal suivant la revendication 12, caractérisée en ce que l'agent réducteur de dureté est présent en une  cipal according to claim 12, characterized in that the hardness reducing agent is present in a quantité correspondant à 5-50 unités d'activité régu-  quantity corresponding to 5-50 units of regular activity lière de Cx/g de la-composition détergente de lavage principal ou à 3-30 unités d'activité modifiée de  Cx / g of the main wash detergent composition or at 3-30 units of modified Cx/g de la composition détergente de lavage principal.  Cx / g of the main wash detergent composition. 14. Composition détergente de lavage prin-  14. Washing detergent composition primarily cipal suivant l'une quelconque des revendications 11  in accordance with one of the claims 11 à 13, caractérisée en ce qu'elle contient également  13, characterized in that it also contains une protéinase bactérienne.bacterial proteinase. 15. Composition détergente de lavage prin-  15. Washing detergent composition primarily cipal suivant la revendication 14, caractérisée en ce que la protéinase bactérienne est une protéinase  cipal according to claim 14, characterized in that the bacterial proteinase is a proteinase obtenue au moyen de Bacillus licheniformis.  obtained with Bacillus licheniformis. 16. Composition détergente de lavage prin-  16. Washing detergent composition primarily cipal suivant l'une quelconque des revendications  in accordance with one of the claims 11 à 15, caractérisée en ce qu'un des ingrédients  11 to 15, characterized in that one of the ingredients détergents est un perborate.detergents is a perborate. 17. Procédé de lavage principal, caracté-  17. Main washing process, characteristics risé en ce que, comme détergent, on utilise un déter-  in that, as a detergent, a detergent is gent de lavage principal suivant l'une quelconque  main wash according to any one of des revendications 11 à 16.Claims 11 to 16. 18. Procédé de lavage principal suivant la revendication 17, caractérisé en ce qu'on utilise la cellulase fongique dans la solution de lavage en une  18. Main washing process according to claim 17, characterized in that the fungal cellulase is used in the washing solution in one concentration correspondant à 10-100 unités dlacti-  concentration corresponding to 10-100 units of vité régulière de Cx/litre de solution de lavage ou  regular consumption of Cx / liter of washing solution or à 6-60 unités d'activité modifiée de Cx/litre de so-  at 6-60 units of modified activity of Cx / liter of lution de lavage.washing solution. 19. Procédé de lavage principal suivant la revendication 18, caractérisé en ce qu'on utilise la cellulase fongique dans la solution de lavage en une  19. Main washing process according to claim 18, characterized in that the fungal cellulase is used in the washing solution in one concentration correspondant à 20-50 unités dlactivi-  concentration corresponding to 20-50 units of activity té régulière de Cx/litre de solution de lavage ou à  regular tee of Cx / liter of wash solution or 12-30 unités d'activité modifiée de Cx/litre de solu-  12-30 units of modified activity of Cx / liter of solu- 248 1712248 1712 tion de lavage.washing. 20. Procédé de lavage principal suivant  20. Main washing process next l'une quelconque des revendications 17 à 19, caracté-  any one of claims 17 to 19, characterized risé en ce que le pH de la solution de lavage se situe entre 7 et 10, de préférence, entre 7,5 et 9,5. 21. Procédé de lavage principal suivant  in that the pH of the wash solution is between 7 and 10, preferably between 7.5 and 9.5. 21. Main washing process next l'une quelconque des revendications 17 à 20, caracté-  any one of claims 17 to 20, characterized risé en ce que la majeure partie du lavage principal  rised in that most of the main wash a lieu à une température inférieure à 700C.  takes place at a temperature below 700C. 22. Procédé de lavage principal suivant la revendication 21, caractérisé en ce que la majeure partie du lavage principal a lieu à une température  22. Main washing process according to claim 21, characterized in that the major part of the main wash takes place at a temperature inférieure à 600C.less than 600C.
FR8108602A 1980-04-30 1981-04-29 ENZYMATIC REDUCING AGENT FOR DETERGENT COMPOSITION Granted FR2481712A1 (en)

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US4435307A (en) 1984-03-06
DE3117250A1 (en) 1982-04-01
MX158929A (en) 1989-03-31
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GB2075028B (en) 1984-03-28
HU196450B (en) 1988-11-28
NL8102123A (en) 1981-11-16
ES501738A0 (en) 1983-06-01
JPS5723699A (en) 1982-02-06
DK187280A (en) 1981-10-31
BR8102643A (en) 1982-01-26
ES8306789A1 (en) 1983-06-01
CH663511GA3 (en) 1987-12-31
FR2481712B1 (en) 1985-04-05
AT384442B (en) 1987-11-10
IT1210475B (en) 1989-09-14
GB2075028A (en) 1981-11-11
IT8121432A0 (en) 1981-04-29
JPS6116316B2 (en) 1986-04-30
NL192043B (en) 1996-09-02
ATA191581A (en) 1987-04-15
BE888632A (en) 1981-10-29

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